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Kinetics of the B-cell population in patients treated with repeated dosage of 100 mg <t>rituximab</t>
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1) Product Images from "Responsiveness to reduced dosage of rituximab in Chinese patients with neuromyelitis optica"

Article Title: Responsiveness to reduced dosage of rituximab in Chinese patients with neuromyelitis optica

Journal: Neurology

doi: 10.1212/WNL.0b013e3182a1aac7

Kinetics of the B-cell population in patients treated with repeated dosage of 100 mg rituximab
Figure Legend Snippet: Kinetics of the B-cell population in patients treated with repeated dosage of 100 mg rituximab

Techniques Used:

Brain and spinal cord MRIs of a patient treated with repeated dosages of 100 mg rituximab
Figure Legend Snippet: Brain and spinal cord MRIs of a patient treated with repeated dosages of 100 mg rituximab

Techniques Used:

2) Product Images from "Epitope interactions of monoclonal antibodies targeting CD20 and their relationship to functional properties"

Article Title: Epitope interactions of monoclonal antibodies targeting CD20 and their relationship to functional properties

Journal: mAbs

doi: 10.4161/mabs.22771

Figure 1. (A) The structure and topology of CD20 and the epitopes recognized by rituximab, ofatumumab and GA101. (B) Sequence alignment of CD20 epitopes recognized by CD20 antibodies based on published information. Core epitope residues are boxed
Figure Legend Snippet: Figure 1. (A) The structure and topology of CD20 and the epitopes recognized by rituximab, ofatumumab and GA101. (B) Sequence alignment of CD20 epitopes recognized by CD20 antibodies based on published information. Core epitope residues are boxed

Techniques Used: Sequencing

Figure 6. Three-dimensional models of A) rituximab and B) GA101. GA101 binds to the same binding epitope region of CD20 as rituximab, but in a different binding orientation. The molecular models were created by combining known structural data
Figure Legend Snippet: Figure 6. Three-dimensional models of A) rituximab and B) GA101. GA101 binds to the same binding epitope region of CD20 as rituximab, but in a different binding orientation. The molecular models were created by combining known structural data

Techniques Used: Binding Assay

Figure 5. Comparison of A) rituximab (Type I) and B) GA101 (Type II) crystal structures in complex with CD20 peptide.29 While for rituximab N171 is deeply immersed and N176 has no contacts with the rituximab CDRs, N171 is not deeply immersed in
Figure Legend Snippet: Figure 5. Comparison of A) rituximab (Type I) and B) GA101 (Type II) crystal structures in complex with CD20 peptide.29 While for rituximab N171 is deeply immersed and N176 has no contacts with the rituximab CDRs, N171 is not deeply immersed in

Techniques Used:

Figure 4. Published crystal structures of CD20 antibodies. A) rituximab-CD20 complex,48 B) ofatumumab (no co-crystal structure is available),50 C) 2H7-CD20 complex,49 and D) GA101-CD20 complex.29 The heavy chain is colored in darker shades, the
Figure Legend Snippet: Figure 4. Published crystal structures of CD20 antibodies. A) rituximab-CD20 complex,48 B) ofatumumab (no co-crystal structure is available),50 C) 2H7-CD20 complex,49 and D) GA101-CD20 complex.29 The heavy chain is colored in darker shades, the

Techniques Used:

Figure 3. Hypothetical model for CD20 binding of Type I and Type II CD20 antibodies explaining the impact of FcγRIIb on internalization. A) Type I antibodies such as rituximab may bind to CD20 in a conformation that allows simultaneous
Figure Legend Snippet: Figure 3. Hypothetical model for CD20 binding of Type I and Type II CD20 antibodies explaining the impact of FcγRIIb on internalization. A) Type I antibodies such as rituximab may bind to CD20 in a conformation that allows simultaneous

Techniques Used: Binding Assay

3) Product Images from "Polyclonal and monoclonal antibodies for treating acute rejection episodes in kidney transplant recipients"

Article Title: Polyclonal and monoclonal antibodies for treating acute rejection episodes in kidney transplant recipients

Journal: The Cochrane Database of Systematic Reviews

doi: 10.1002/14651858.CD004756.pub4

Comparison 4 Treatment of first rejection (B cell): rituximab + steroids versus steroids alone, Outcome 3 Graft loss or death with a functioning graft within 12 months.
Figure Legend Snippet: Comparison 4 Treatment of first rejection (B cell): rituximab + steroids versus steroids alone, Outcome 3 Graft loss or death with a functioning graft within 12 months.

Techniques Used:

Comparison 4 Treatment of first rejection (B cell): rituximab + steroids versus steroids alone, Outcome 4 Death within 12 months.
Figure Legend Snippet: Comparison 4 Treatment of first rejection (B cell): rituximab + steroids versus steroids alone, Outcome 4 Death within 12 months.

Techniques Used:

Comparison 4 Treatment of first rejection (B cell): rituximab + steroids versus steroids alone, Outcome 2 Additional treatment required.
Figure Legend Snippet: Comparison 4 Treatment of first rejection (B cell): rituximab + steroids versus steroids alone, Outcome 2 Additional treatment required.

Techniques Used:

Comparison 4 Treatment of first rejection (B cell): rituximab + steroids versus steroids alone, Outcome 1 Failure of reversal of acute rejection.
Figure Legend Snippet: Comparison 4 Treatment of first rejection (B cell): rituximab + steroids versus steroids alone, Outcome 1 Failure of reversal of acute rejection.

Techniques Used:

Comparison 4 Treatment of first rejection (B cell): rituximab + steroids versus steroids alone, Outcome 5 Treatment adverse events.
Figure Legend Snippet: Comparison 4 Treatment of first rejection (B cell): rituximab + steroids versus steroids alone, Outcome 5 Treatment adverse events.

Techniques Used:

4) Product Images from "A Novel Hemolytic Complement-Sufficient NSG Mouse Model Supports Studies of Complement-Mediated Antitumor Activity In Vivo"

Article Title: A Novel Hemolytic Complement-Sufficient NSG Mouse Model Supports Studies of Complement-Mediated Antitumor Activity In Vivo

Journal: Journal of immunological methods

doi: 10.1016/j.jim.2017.03.021

(A) Change in body weight over time in different cohorts of age- and sex-matched NSG and NSG-Hc 1 mice engrafted with Daudi cells. (B) Kaplan-Meier survival plot demonstrate percent of survival over time in different cohorts of age- and sex-matched NSG and NSG-Hc 1 mice engrafted with Daudi cells. Non-engrafted NSG mice were used as negative controls. Data are reported as percentage of the mean ± SE. PBS, phosphate buffer saline; RTX, rituximab; p
Figure Legend Snippet: (A) Change in body weight over time in different cohorts of age- and sex-matched NSG and NSG-Hc 1 mice engrafted with Daudi cells. (B) Kaplan-Meier survival plot demonstrate percent of survival over time in different cohorts of age- and sex-matched NSG and NSG-Hc 1 mice engrafted with Daudi cells. Non-engrafted NSG mice were used as negative controls. Data are reported as percentage of the mean ± SE. PBS, phosphate buffer saline; RTX, rituximab; p

Techniques Used: Mouse Assay

Representative flow cytometry analysis of human CD45+ cells in (A) peripheral blood and (B) bone marrow 38 days post-engraftment demonstrating percentages of viable cells in different cohorts of age- and sex-matched NSG and NSG-Hc 1 mice displaying levels of Daudi cell engraftment. Non-engrafted NSG mice were used as negative controls (data not shown). Data are reported as percentage of the mean ± SD. PBS, phosphate buffer saline; RTX, rituximab; p
Figure Legend Snippet: Representative flow cytometry analysis of human CD45+ cells in (A) peripheral blood and (B) bone marrow 38 days post-engraftment demonstrating percentages of viable cells in different cohorts of age- and sex-matched NSG and NSG-Hc 1 mice displaying levels of Daudi cell engraftment. Non-engrafted NSG mice were used as negative controls (data not shown). Data are reported as percentage of the mean ± SD. PBS, phosphate buffer saline; RTX, rituximab; p

Techniques Used: Flow Cytometry, Cytometry, Mouse Assay

Representative images of the (A) ovary and (B) kidney 38 days post-engraftment displaying level of Daudi cell engraftment (brown stained cells) in different cohorts of age- and sex-matched rituximab-treated NSG and NSG- Hc 1 mice. Low-magnification images at top right showing anti-CD45 staining IHC, at bottom right showing H E staining, (×10, scale bar = 200um) showing; high-magnification image (×20, scale bar = 100um) showing anti-human CD45 IHC. PBS, phosphate buffer saline; RTX, rituximab. There were also increased numbers of Daudi cells in the livers, spleens, lungs, and lymph nodes of NSG mice (data not shown).
Figure Legend Snippet: Representative images of the (A) ovary and (B) kidney 38 days post-engraftment displaying level of Daudi cell engraftment (brown stained cells) in different cohorts of age- and sex-matched rituximab-treated NSG and NSG- Hc 1 mice. Low-magnification images at top right showing anti-CD45 staining IHC, at bottom right showing H E staining, (×10, scale bar = 200um) showing; high-magnification image (×20, scale bar = 100um) showing anti-human CD45 IHC. PBS, phosphate buffer saline; RTX, rituximab. There were also increased numbers of Daudi cells in the livers, spleens, lungs, and lymph nodes of NSG mice (data not shown).

Techniques Used: Staining, Mouse Assay, Immunohistochemistry

5) Product Images from "Ofatumumab Exhibits Enhanced In Vitro and In Vivo Activity Compared to Rituximab in Preclinical Models of Mantle Cell Lymphoma"

Article Title: Ofatumumab Exhibits Enhanced In Vitro and In Vivo Activity Compared to Rituximab in Preclinical Models of Mantle Cell Lymphoma

Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

doi: 10.1158/1078-0432.CCR-15-0056

Expression of CD20, CD55, and CD59 in MCL cell lines compared with rituximab-sensitive Raji and rituximab-resistant Raji 4RH cell lines. The surface density of CD20, CD55, and CD59 was determined using Imagestream. MCL cell lines exhibit similar density of CD20 expression on their cell membrane when compared with the rituximab-sensitive Raji cell line. However, the complement inhibitory proteins CD55 and CD59 are present at increased surface density compared with Raji cells, similar to the rituximab-resistant Raji 4RH cell line.
Figure Legend Snippet: Expression of CD20, CD55, and CD59 in MCL cell lines compared with rituximab-sensitive Raji and rituximab-resistant Raji 4RH cell lines. The surface density of CD20, CD55, and CD59 was determined using Imagestream. MCL cell lines exhibit similar density of CD20 expression on their cell membrane when compared with the rituximab-sensitive Raji cell line. However, the complement inhibitory proteins CD55 and CD59 are present at increased surface density compared with Raji cells, similar to the rituximab-resistant Raji 4RH cell line.

Techniques Used: Expressing

Ofatumumab and rituximab induce a similar degree of ADCC-associated cell lyis. MCL tumor cell lines were tested for ADCC cell lysis using a 51 Cr release assay using effector cells from normal healthy donors at a ratio of 40:1. In all MCL cell lines tested, ofatumumab and rituximab induced similar levels of ADCC lysis at a dose of 10 μg/mL.
Figure Legend Snippet: Ofatumumab and rituximab induce a similar degree of ADCC-associated cell lyis. MCL tumor cell lines were tested for ADCC cell lysis using a 51 Cr release assay using effector cells from normal healthy donors at a ratio of 40:1. In all MCL cell lines tested, ofatumumab and rituximab induced similar levels of ADCC lysis at a dose of 10 μg/mL.

Techniques Used: Lysis, Release Assay

In MCL cells derived from patient samples, ofatumumab induced similar or more cell lysis by CDC compared with rituximab. B cells were isolated from pretreatment tumor samples ( n = 4) obtained from MCL patients and tested for monoclonal antibody response by CellTiter Glo assay using rituximab or ofatumumab at a concentration of 10 μg/mL in the presence of human serum as a source of complement. Two of the four samples exhibit a statistically significant decrease in viability following ofatumumab exposure as compared to rituximab. The remaining samples show a similar response to each antibody.
Figure Legend Snippet: In MCL cells derived from patient samples, ofatumumab induced similar or more cell lysis by CDC compared with rituximab. B cells were isolated from pretreatment tumor samples ( n = 4) obtained from MCL patients and tested for monoclonal antibody response by CellTiter Glo assay using rituximab or ofatumumab at a concentration of 10 μg/mL in the presence of human serum as a source of complement. Two of the four samples exhibit a statistically significant decrease in viability following ofatumumab exposure as compared to rituximab. The remaining samples show a similar response to each antibody.

Techniques Used: Derivative Assay, Lysis, Isolation, Glo Assay, Concentration Assay

Ofatumumab and rituximab exhibit modest direct antitumor activity against MCL cell lines. MCL cell lines were utilized in alamarBlue reduction assays to investigate the direct antilymphoma activity of each antibody. Each antibody exhibited a time-dependent decrease in MCL cell viability following exposure to 10 μg/mL of each respective antibody in the presence of a cross-linking IgG antibody. With the exception of the Mino cell line at the 72-hour time point, all cell lines demonstrated similar degrees of direct antitumor activity with ofatumumab and rituximab at the dose tested (*, P
Figure Legend Snippet: Ofatumumab and rituximab exhibit modest direct antitumor activity against MCL cell lines. MCL cell lines were utilized in alamarBlue reduction assays to investigate the direct antilymphoma activity of each antibody. Each antibody exhibited a time-dependent decrease in MCL cell viability following exposure to 10 μg/mL of each respective antibody in the presence of a cross-linking IgG antibody. With the exception of the Mino cell line at the 72-hour time point, all cell lines demonstrated similar degrees of direct antitumor activity with ofatumumab and rituximab at the dose tested (*, P

Techniques Used: Activity Assay

Ofatumumab induces significantly higher levels of CDC-associated cell lysis than rituximab. MCL tumor cell lines were tested for CDC cell lysis using a 51 Cr release assay with human serum from normal healthy donors as a source of complement. In all MCL cell lines tested, with the exception of Granta, ofatumumab was more effective than rituximab at inducing CDC lysis at a dose of 10 μg/mL. (*, P
Figure Legend Snippet: Ofatumumab induces significantly higher levels of CDC-associated cell lysis than rituximab. MCL tumor cell lines were tested for CDC cell lysis using a 51 Cr release assay with human serum from normal healthy donors as a source of complement. In all MCL cell lines tested, with the exception of Granta, ofatumumab was more effective than rituximab at inducing CDC lysis at a dose of 10 μg/mL. (*, P

Techniques Used: Lysis, Release Assay

6) Product Images from "Update on Hairy Cell Leukemia"

Article Title: Update on Hairy Cell Leukemia

Journal: Clinical advances in hematology & oncology : H & O

doi:

The schematic structures of anti–hairy cell leukemia agents. The traditional purine analogues pentostatin and cladribine have similar structures but slightly different mechanisms of action. Bendamustine contains the benzimidazole ring of cladribine, which makes it a purine analogue, but has alkylating groups for additional mechanisms of action. LMB-2 contains the variable domains (VH and VL) of the anti-CD25 monoclonal antibody anti-Tac connected by a (G 4 S) 3 peptide linker, with VL fused to PE38. The anti-CD22 recombinant immunotoxins BL22 and moxetumomab pasudotox contain a disulfide-stabilized Fv made by engineering a disulfide bond into the framework regions, replacing Arg44 of VH and Gly100 of VL. In moxetumomab pasudotox, the amino acids 100, 100a, and 100b of VH, in the third complementarity-determining region (CDR3), are mutated from SSY (serine, serine, and tyrosine) to THW (threonine, histidine, and tryptophan), resulting in 14-fold improvement in binding affinity. Rituximab is a chimeric monoclonal antibody containing human CH3 and CH4 constant domains, whereas the remainder is of murine origin. The structures of the oral small molecules ibrutinib, vemurafenib, dabrafenib, and trametinib are shown. CDA, 2-chlorodeoxyadenosine; DCF, 2-deoxycoformycin; Fv, variable fragment.
Figure Legend Snippet: The schematic structures of anti–hairy cell leukemia agents. The traditional purine analogues pentostatin and cladribine have similar structures but slightly different mechanisms of action. Bendamustine contains the benzimidazole ring of cladribine, which makes it a purine analogue, but has alkylating groups for additional mechanisms of action. LMB-2 contains the variable domains (VH and VL) of the anti-CD25 monoclonal antibody anti-Tac connected by a (G 4 S) 3 peptide linker, with VL fused to PE38. The anti-CD22 recombinant immunotoxins BL22 and moxetumomab pasudotox contain a disulfide-stabilized Fv made by engineering a disulfide bond into the framework regions, replacing Arg44 of VH and Gly100 of VL. In moxetumomab pasudotox, the amino acids 100, 100a, and 100b of VH, in the third complementarity-determining region (CDR3), are mutated from SSY (serine, serine, and tyrosine) to THW (threonine, histidine, and tryptophan), resulting in 14-fold improvement in binding affinity. Rituximab is a chimeric monoclonal antibody containing human CH3 and CH4 constant domains, whereas the remainder is of murine origin. The structures of the oral small molecules ibrutinib, vemurafenib, dabrafenib, and trametinib are shown. CDA, 2-chlorodeoxyadenosine; DCF, 2-deoxycoformycin; Fv, variable fragment.

Techniques Used: Recombinant, Binding Assay

Treatment algorithm for HCL and HCLv. Both standard and investigational treatment options are shown for (1) patients with HCL without prior purine analogue therapy, (2) patients with HCL treated with 1 prior purine analogue, (3) patients with HCLv treated with 0 or 1 prior purine analogue, and (4) patients with multiply relapsed HCL/HCLv. BR, bendamustine/rituximab; CDA, cladribine (2-chlorodeoxyadenosine); CDAR, cladribine/rituximab; DCF, pentostatin (2’-deoxycoformycin); DCFR, pentostatin/rituximab; HCL, hairy cell leukemia; HCLv, hairy cell leukemia variant; tx, therapy.
Figure Legend Snippet: Treatment algorithm for HCL and HCLv. Both standard and investigational treatment options are shown for (1) patients with HCL without prior purine analogue therapy, (2) patients with HCL treated with 1 prior purine analogue, (3) patients with HCLv treated with 0 or 1 prior purine analogue, and (4) patients with multiply relapsed HCL/HCLv. BR, bendamustine/rituximab; CDA, cladribine (2-chlorodeoxyadenosine); CDAR, cladribine/rituximab; DCF, pentostatin (2’-deoxycoformycin); DCFR, pentostatin/rituximab; HCL, hairy cell leukemia; HCLv, hairy cell leukemia variant; tx, therapy.

Techniques Used: Variant Assay

7) Product Images from "NK-cell activation and antibody-dependent cellular cytotoxicity induced by rituximab-coated target cells is inhibited by the C3b component of complement"

Article Title: NK-cell activation and antibody-dependent cellular cytotoxicity induced by rituximab-coated target cells is inhibited by the C3b component of complement

Journal: Blood

doi: 10.1182/blood-2007-02-074716

Serum-depleted C5 inhibits NK-cell CD54 up-regulation, while serum-depleted C1q or C3 does not. PBMCs were mixed with Raji cells at a 1:1 ratio for 20 hours with 5 μg/mL of rituximab in the presence or absence of C1q-, C3-, or C5-depleted serum. Surface marker expression was determined using flow cytometry with gating on CD3 − , CD56 + lymphocytes. NK-cell CD54, expressed as a percentage of CD54 bright, was determined. (A) Evaluation in C1q-depleted serum, C1q-depleted serum with purified C1q added back, or purified C1q alone (n = 3). (B) Evaluation in C3-depleted serum, C3-depleted serum with purified C3 added back, or purified C3 alone (n = 3). (C) Evaluation in various concentrations of C5-depleted serum (n = 3). Error bars represent SD of the mean.
Figure Legend Snippet: Serum-depleted C5 inhibits NK-cell CD54 up-regulation, while serum-depleted C1q or C3 does not. PBMCs were mixed with Raji cells at a 1:1 ratio for 20 hours with 5 μg/mL of rituximab in the presence or absence of C1q-, C3-, or C5-depleted serum. Surface marker expression was determined using flow cytometry with gating on CD3 − , CD56 + lymphocytes. NK-cell CD54, expressed as a percentage of CD54 bright, was determined. (A) Evaluation in C1q-depleted serum, C1q-depleted serum with purified C1q added back, or purified C1q alone (n = 3). (B) Evaluation in C3-depleted serum, C3-depleted serum with purified C3 added back, or purified C3 alone (n = 3). (C) Evaluation in various concentrations of C5-depleted serum (n = 3). Error bars represent SD of the mean.

Techniques Used: Marker, Expressing, Flow Cytometry, Cytometry, Purification

C5-depleted serum inhibits rituximab-mediated ADCC. Raji cells were labeled with 51 Cr and incubated with purified NK cells in various E/T ratios, 5 μg/mL rituximab, and either C5-depleted serum, heat-inactivated C5-depleted serum, or media. Percentage of specific lysis was measured based on 51 Cr release. All samples were run in triplicate (n = 3). Data are representative of 3 independent experiments. Error bars represent SD of the mean.
Figure Legend Snippet: C5-depleted serum inhibits rituximab-mediated ADCC. Raji cells were labeled with 51 Cr and incubated with purified NK cells in various E/T ratios, 5 μg/mL rituximab, and either C5-depleted serum, heat-inactivated C5-depleted serum, or media. Percentage of specific lysis was measured based on 51 Cr release. All samples were run in triplicate (n = 3). Data are representative of 3 independent experiments. Error bars represent SD of the mean.

Techniques Used: Labeling, Incubation, Purification, Lysis

NK cell CD16 binding to rituximab is blocked by complement fixation. U-bottomed 96-well microtiter plates were coated with varying concentrations of rituximab. Wells were incubated with media, 50% serum, or 50% heat-inactivated serum. After washing, NK cells were allowed to sit on plate for 30 minutes at room temperature. High absorbance resulted from NK-cell pelleting that occurred in the absence of binding to rituximab. All samples were run in triplicate (n = 3). Data are representative of 3 independent experiments. Error bars represent SD of the mean.
Figure Legend Snippet: NK cell CD16 binding to rituximab is blocked by complement fixation. U-bottomed 96-well microtiter plates were coated with varying concentrations of rituximab. Wells were incubated with media, 50% serum, or 50% heat-inactivated serum. After washing, NK cells were allowed to sit on plate for 30 minutes at room temperature. High absorbance resulted from NK-cell pelleting that occurred in the absence of binding to rituximab. All samples were run in triplicate (n = 3). Data are representative of 3 independent experiments. Error bars represent SD of the mean.

Techniques Used: Binding Assay, Incubation

Inhibitory effect of serum is dose dependent and abrogated by heat inactivation. PBMCs and Raji cells were mixed at a 1:1 ratio for 20 hours in varying concentrations of serum or heat-inactivated serum in the presence of 0.2 μg/mL rituximab. Surface marker expression was determined using flow cytometry with gating on CD3 − , CD56 + lymphocytes. (A) NK-cell CD16, expressed as median fluorescence, after incubation with varying concentrations of serum and heat-inactivated serum (n = 3). (B) NK-cell CD54, expressed as a percentage of CD54 bright, after incubation with varying concentrations of serum and heat inactivated serum (n = 3). Error bars represent SD of the mean.
Figure Legend Snippet: Inhibitory effect of serum is dose dependent and abrogated by heat inactivation. PBMCs and Raji cells were mixed at a 1:1 ratio for 20 hours in varying concentrations of serum or heat-inactivated serum in the presence of 0.2 μg/mL rituximab. Surface marker expression was determined using flow cytometry with gating on CD3 − , CD56 + lymphocytes. (A) NK-cell CD16, expressed as median fluorescence, after incubation with varying concentrations of serum and heat-inactivated serum (n = 3). (B) NK-cell CD54, expressed as a percentage of CD54 bright, after incubation with varying concentrations of serum and heat inactivated serum (n = 3). Error bars represent SD of the mean.

Techniques Used: Marker, Expressing, Flow Cytometry, Cytometry, Fluorescence, Incubation

Target cells are lysed in the presence of serum and rituximab irrespective of presence of effector cells. Raji cells were incubated for 20 hours in media, 50% serum, or 50% serum plus PBMCs with varying concentrations of rituximab. The percent of viable target cells were determined using flow cytometry by counting annexin V and propidium iodide–negative target cells (n = 3). Error bars represent SD of the mean.
Figure Legend Snippet: Target cells are lysed in the presence of serum and rituximab irrespective of presence of effector cells. Raji cells were incubated for 20 hours in media, 50% serum, or 50% serum plus PBMCs with varying concentrations of rituximab. The percent of viable target cells were determined using flow cytometry by counting annexin V and propidium iodide–negative target cells (n = 3). Error bars represent SD of the mean.

Techniques Used: Incubation, Flow Cytometry, Cytometry

Serum inhibits changes in CD16 expression in the absence of viable target cells. (A) Raji cells were fixed in 1% formaldehyde and washed extensively. Fixed cells were mixed with PBMCs at a 1:1 ratio for 20 hours with 5 μg/mL rituximab (n = 3). (B) Flat-bottom plates were coated with 10 μg/mL of rituximab. Plates were washed, and PBMCs were added and cultured for 20 hours. Incubations with fixed Raji cells and rituximab or rituximab-coated plates were preformed in the presence or absence of 50% serum and heat-inactivated serum (n = 5). Surface marker expression was determined using flow cytometry with gating on CD3 − , CD56 + lymphocytes. NK-cell CD16, expressed as median fluorescence, was measured after incubation with media, serum, and heat-inactivated serum in the presence of fixed Raji cells (A) or flat-bottom plates coated with rituximab (B). Error bars represent SD of the mean.
Figure Legend Snippet: Serum inhibits changes in CD16 expression in the absence of viable target cells. (A) Raji cells were fixed in 1% formaldehyde and washed extensively. Fixed cells were mixed with PBMCs at a 1:1 ratio for 20 hours with 5 μg/mL rituximab (n = 3). (B) Flat-bottom plates were coated with 10 μg/mL of rituximab. Plates were washed, and PBMCs were added and cultured for 20 hours. Incubations with fixed Raji cells and rituximab or rituximab-coated plates were preformed in the presence or absence of 50% serum and heat-inactivated serum (n = 5). Surface marker expression was determined using flow cytometry with gating on CD3 − , CD56 + lymphocytes. NK-cell CD16, expressed as median fluorescence, was measured after incubation with media, serum, and heat-inactivated serum in the presence of fixed Raji cells (A) or flat-bottom plates coated with rituximab (B). Error bars represent SD of the mean.

Techniques Used: Expressing, Cell Culture, Marker, Flow Cytometry, Cytometry, Fluorescence, Incubation

C3b-stabilizing antibody (3E7) enhances the inhibition of NK-cell CD16 down-modulation and CD54 up-regulation. PBMCs and Raji cells were mixed at a 1:1 ratio for 20 hours in the presence or absence of 50% serum with varying concentrations of rituximab and 10 μg/mL of 3E7. Surface marker expression was determined using flow cytometry with gating on CD3 − , CD56 + lymphocytes. NK-cell CD54, expressed as a percentage of CD54 bright, was cultured in media, serum, or serum plus 3E7 (n = 3). Error bars represent SD of the mean.
Figure Legend Snippet: C3b-stabilizing antibody (3E7) enhances the inhibition of NK-cell CD16 down-modulation and CD54 up-regulation. PBMCs and Raji cells were mixed at a 1:1 ratio for 20 hours in the presence or absence of 50% serum with varying concentrations of rituximab and 10 μg/mL of 3E7. Surface marker expression was determined using flow cytometry with gating on CD3 − , CD56 + lymphocytes. NK-cell CD54, expressed as a percentage of CD54 bright, was cultured in media, serum, or serum plus 3E7 (n = 3). Error bars represent SD of the mean.

Techniques Used: Inhibition, Marker, Expressing, Flow Cytometry, Cytometry, Cell Culture

Serum inhibits rituximab-induced NK-cell CD16 down-modulation and CD54 up-regulation. PBMCs and Raji, Daudi, or FL cells were mixed at a 1:1 ratio for 20 hours in the presence or absence of 50% serum with varying concentrations of rituximab. Surface marker expression was determined using flow cytometry with gating on CD3 − , CD56 + lymphocytes. (A) NK-cell CD16, expressed as median fluorescence, in the absence and presence of serum (n = 3). (B) NK-cell CD54, expressed as a percentage of CD54 bright, in the absence and presence of serum (n = 3). Error bars represent the standard deviation (SD) of the mean.
Figure Legend Snippet: Serum inhibits rituximab-induced NK-cell CD16 down-modulation and CD54 up-regulation. PBMCs and Raji, Daudi, or FL cells were mixed at a 1:1 ratio for 20 hours in the presence or absence of 50% serum with varying concentrations of rituximab. Surface marker expression was determined using flow cytometry with gating on CD3 − , CD56 + lymphocytes. (A) NK-cell CD16, expressed as median fluorescence, in the absence and presence of serum (n = 3). (B) NK-cell CD54, expressed as a percentage of CD54 bright, in the absence and presence of serum (n = 3). Error bars represent the standard deviation (SD) of the mean.

Techniques Used: Marker, Expressing, Flow Cytometry, Cytometry, Fluorescence, Standard Deviation

8) Product Images from "Application of a novel inhibitor of human CD59 for the enhancement of complement-dependent cytolysis on cancer cells"

Article Title: Application of a novel inhibitor of human CD59 for the enhancement of complement-dependent cytolysis on cancer cells

Journal: Cellular and Molecular Immunology

doi: 10.1038/cmi.2010.35

The resistance of the ARH-77 cells to the rituximab-mediated CDC effect is associated with CD59 and other mCRPs levels. ( a ) CD46 and CD55 are expressed on the surface of RL-7, RR51.2 and ARH-77 cells. Shaded histogram: isotype-matched Abs+FITC
Figure Legend Snippet: The resistance of the ARH-77 cells to the rituximab-mediated CDC effect is associated with CD59 and other mCRPs levels. ( a ) CD46 and CD55 are expressed on the surface of RL-7, RR51.2 and ARH-77 cells. Shaded histogram: isotype-matched Abs+FITC

Techniques Used:

rILYd4 sensitized the original RL-7 lymphoma cells and rituximab-induced resistant RR51.2 cells to the rituximab-mediated CDC effect. ( a , b ) RL-7 cells and RR51.2 cells expressed CD20 and CD59 on the cell surface. Shaded histogram: isotype-matched Abs+FITC
Figure Legend Snippet: rILYd4 sensitized the original RL-7 lymphoma cells and rituximab-induced resistant RR51.2 cells to the rituximab-mediated CDC effect. ( a , b ) RL-7 cells and RR51.2 cells expressed CD20 and CD59 on the cell surface. Shaded histogram: isotype-matched Abs+FITC

Techniques Used:

rILYd4 senstized MM ARH-77 cells to rituximab-mediated CDC effect. ( a ) CD20 and CD59 are expressed on the surface of MM ARH-77 cells. Shaded histogram: isotype-matched Abs+FITC secondary Ab staining (negative control); open histogram: anti-hCD59
Figure Legend Snippet: rILYd4 senstized MM ARH-77 cells to rituximab-mediated CDC effect. ( a ) CD20 and CD59 are expressed on the surface of MM ARH-77 cells. Shaded histogram: isotype-matched Abs+FITC secondary Ab staining (negative control); open histogram: anti-hCD59

Techniques Used: Staining, Negative Control

9) Product Images from "Polyclonal and monoclonal antibodies for treating acute rejection episodes in kidney transplant recipients"

Article Title: Polyclonal and monoclonal antibodies for treating acute rejection episodes in kidney transplant recipients

Journal: The Cochrane Database of Systematic Reviews

doi: 10.1002/14651858.CD004756.pub4

Comparison 4 Treatment of first rejection (B cell): rituximab + steroids versus steroids alone, Outcome 3 Graft loss or death with a functioning graft within 12 months.
Figure Legend Snippet: Comparison 4 Treatment of first rejection (B cell): rituximab + steroids versus steroids alone, Outcome 3 Graft loss or death with a functioning graft within 12 months.

Techniques Used:

Comparison 4 Treatment of first rejection (B cell): rituximab + steroids versus steroids alone, Outcome 4 Death within 12 months.
Figure Legend Snippet: Comparison 4 Treatment of first rejection (B cell): rituximab + steroids versus steroids alone, Outcome 4 Death within 12 months.

Techniques Used:

Comparison 4 Treatment of first rejection (B cell): rituximab + steroids versus steroids alone, Outcome 2 Additional treatment required.
Figure Legend Snippet: Comparison 4 Treatment of first rejection (B cell): rituximab + steroids versus steroids alone, Outcome 2 Additional treatment required.

Techniques Used:

Comparison 4 Treatment of first rejection (B cell): rituximab + steroids versus steroids alone, Outcome 1 Failure of reversal of acute rejection.
Figure Legend Snippet: Comparison 4 Treatment of first rejection (B cell): rituximab + steroids versus steroids alone, Outcome 1 Failure of reversal of acute rejection.

Techniques Used:

Comparison 4 Treatment of first rejection (B cell): rituximab + steroids versus steroids alone, Outcome 5 Treatment adverse events.
Figure Legend Snippet: Comparison 4 Treatment of first rejection (B cell): rituximab + steroids versus steroids alone, Outcome 5 Treatment adverse events.

Techniques Used:

10) Product Images from "Depletion of the C3 component of complement enhances the ability of rituximab-coated target cells to activate human NK cells and improves the efficacy of monoclonal antibody therapy in an in vivo model"

Article Title: Depletion of the C3 component of complement enhances the ability of rituximab-coated target cells to activate human NK cells and improves the efficacy of monoclonal antibody therapy in an in vivo model

Journal: Blood

doi: 10.1182/blood-2009-01-200469

Transudative pleural fluid and nonmalignant ascites inhibit rituximab-induced NK cell CD16 down-modulation and CD54 up-regulation . PBMCs and Raji cells were mixed at a 1:1 ratio for 20 hours in the presence or absence of 50% pleural fluid or ascites with
Figure Legend Snippet: Transudative pleural fluid and nonmalignant ascites inhibit rituximab-induced NK cell CD16 down-modulation and CD54 up-regulation . PBMCs and Raji cells were mixed at a 1:1 ratio for 20 hours in the presence or absence of 50% pleural fluid or ascites with

Techniques Used:

CVF enhances NK cell CD54 up-regulation in the presence of complement and rituximab-coated target cells . PBMCs and Raji cells were mixed at a 1:1 ratio for 20 hours in the presence of 50% serum, pleural fluid, or ascites with or without the addition of
Figure Legend Snippet: CVF enhances NK cell CD54 up-regulation in the presence of complement and rituximab-coated target cells . PBMCs and Raji cells were mixed at a 1:1 ratio for 20 hours in the presence of 50% serum, pleural fluid, or ascites with or without the addition of

Techniques Used:

11) Product Images from "An IL-15 superagonist/IL-15Rα fusion complex protects and rescues NK cell cytotoxic function from TGF-β1-mediated immunosuppression"

Article Title: An IL-15 superagonist/IL-15Rα fusion complex protects and rescues NK cell cytotoxic function from TGF-β1-mediated immunosuppression

Journal: Cancer immunology, immunotherapy : CII

doi: 10.1007/s00262-018-2121-4

IL-15SA/IL-15RA inhibited TGF-β1 from suppressing NK cell-ADCC activity. (a) The expression of programmed death-ligand 1 (PD-L1) (solid histograms) and (b) epidermal growth factor receptor (EGFR) (solid histograms) in four tumor cell lines: lung cancer H460, prostate cancer LNCap, estrogen receptor positive breast cancer MCF7, and triple negative breast cancer MDA-MB-231. Dot histograms depict isotype control for each antibody. Inset numbers indicate % positive cells and mean fluorescence intensity (MFI) (parentheses). (c) ADCC assays were performed using three tumor cell lines. NK cells obtained from healthy donors were untreated or treated with TGF-β1 (2 ng/ml) and/or IL-15SA/IL-15RA (50 ng/ml) for 48 hours, then washed and used for ADCC assays at an E:T ratio of 10:1. To mediate ADCC, the following IgG1 mAbs were utilized: rituximab as an isotype control, avelumab as anti-PD-L1 antibody, cetuximab as anti-EGFR antibody. Statistical analyses were done by Student’s t -test, * = p
Figure Legend Snippet: IL-15SA/IL-15RA inhibited TGF-β1 from suppressing NK cell-ADCC activity. (a) The expression of programmed death-ligand 1 (PD-L1) (solid histograms) and (b) epidermal growth factor receptor (EGFR) (solid histograms) in four tumor cell lines: lung cancer H460, prostate cancer LNCap, estrogen receptor positive breast cancer MCF7, and triple negative breast cancer MDA-MB-231. Dot histograms depict isotype control for each antibody. Inset numbers indicate % positive cells and mean fluorescence intensity (MFI) (parentheses). (c) ADCC assays were performed using three tumor cell lines. NK cells obtained from healthy donors were untreated or treated with TGF-β1 (2 ng/ml) and/or IL-15SA/IL-15RA (50 ng/ml) for 48 hours, then washed and used for ADCC assays at an E:T ratio of 10:1. To mediate ADCC, the following IgG1 mAbs were utilized: rituximab as an isotype control, avelumab as anti-PD-L1 antibody, cetuximab as anti-EGFR antibody. Statistical analyses were done by Student’s t -test, * = p

Techniques Used: Activity Assay, Expressing, Multiple Displacement Amplification, Fluorescence

12) Product Images from "Phase 1/2 study of lumiliximab combined with fludarabine, cyclophosphamide, and rituximab in patients with relapsed or refractory chronic lymphocytic leukemia"

Article Title: Phase 1/2 study of lumiliximab combined with fludarabine, cyclophosphamide, and rituximab in patients with relapsed or refractory chronic lymphocytic leukemia

Journal: Blood

doi: 10.1182/blood-2009-08-237727

Plasma concentrations . Mean ± SD plasma concentrations of lumiliximab (A) and rituximab (B) during cycles 1, 3, and 6 of treatment for patients who received 500 mg/m 2 of lumiliximab.
Figure Legend Snippet: Plasma concentrations . Mean ± SD plasma concentrations of lumiliximab (A) and rituximab (B) during cycles 1, 3, and 6 of treatment for patients who received 500 mg/m 2 of lumiliximab.

Techniques Used:

Kaplan-Meier analyses of response . PFS for all patients (A) and DR (B) for all responders (PR and CR) treated with fludarabine, cyclophosphamide, rituximab, and lumiliximab.
Figure Legend Snippet: Kaplan-Meier analyses of response . PFS for all patients (A) and DR (B) for all responders (PR and CR) treated with fludarabine, cyclophosphamide, rituximab, and lumiliximab.

Techniques Used:

13) Product Images from "An Antibody-based Multifaceted Approach Targeting the Human Transferrin Receptor for the Treatment of B-cell Malignancies"

Article Title: An Antibody-based Multifaceted Approach Targeting the Human Transferrin Receptor for the Treatment of B-cell Malignancies

Journal: Journal of immunotherapy (Hagerstown, Md. : 1997)

doi: 10.1097/CJI.0b013e318222ffc8

Antibody-mediated C1q binding on the surface of target cells. IM-9, U266, HL-60, or Ramos cells were incubated with 5 μg/mL of rituximab, the parental ch128.1, or the fusion protein ch128.1Av for 30 minutes at room temperature. Samples treated
Figure Legend Snippet: Antibody-mediated C1q binding on the surface of target cells. IM-9, U266, HL-60, or Ramos cells were incubated with 5 μg/mL of rituximab, the parental ch128.1, or the fusion protein ch128.1Av for 30 minutes at room temperature. Samples treated

Techniques Used: Binding Assay, Incubation

14) Product Images from "Anti-CD20 Agents for Multiple Sclerosis: Spotlight on Ocrelizumab and Ofatumumab"

Article Title: Anti-CD20 Agents for Multiple Sclerosis: Spotlight on Ocrelizumab and Ofatumumab

Journal: Brain Sciences

doi: 10.3390/brainsci10100758

Three Anti-CD20 Antibodies: Rituximab, Ocrelizumab, and Ofatumumab. mAb: monoclonal antibody, Th: T-Helper, IL: Interleukin.
Figure Legend Snippet: Three Anti-CD20 Antibodies: Rituximab, Ocrelizumab, and Ofatumumab. mAb: monoclonal antibody, Th: T-Helper, IL: Interleukin.

Techniques Used:

15) Product Images from "Insights into the effector functions of human IgG3 in the context of an antibody targeting transferrin receptor 1"

Article Title: Insights into the effector functions of human IgG3 in the context of an antibody targeting transferrin receptor 1

Journal: Molecular immunology

doi: 10.1016/j.molimm.2015.07.001

Induction of CDC by ch128.1 and its mutants. Calcein AM-labeled Ramos cells were incubated with 1 μg/mL of the specified antibodies, rituximab positive control, or an anti-human HER2/ neu IgG3/κ as an isotype control. Human complement was
Figure Legend Snippet: Induction of CDC by ch128.1 and its mutants. Calcein AM-labeled Ramos cells were incubated with 1 μg/mL of the specified antibodies, rituximab positive control, or an anti-human HER2/ neu IgG3/κ as an isotype control. Human complement was

Techniques Used: Labeling, Incubation, Positive Control

16) Product Images from "Polyclonal and monoclonal antibodies for treating acute rejection episodes in kidney transplant recipients"

Article Title: Polyclonal and monoclonal antibodies for treating acute rejection episodes in kidney transplant recipients

Journal: The Cochrane Database of Systematic Reviews

doi: 10.1002/14651858.CD004756.pub4

Comparison 4 Treatment of first rejection (B cell): rituximab + steroids versus steroids alone, Outcome 3 Graft loss or death with a functioning graft within 12 months.
Figure Legend Snippet: Comparison 4 Treatment of first rejection (B cell): rituximab + steroids versus steroids alone, Outcome 3 Graft loss or death with a functioning graft within 12 months.

Techniques Used:

Comparison 4 Treatment of first rejection (B cell): rituximab + steroids versus steroids alone, Outcome 4 Death within 12 months.
Figure Legend Snippet: Comparison 4 Treatment of first rejection (B cell): rituximab + steroids versus steroids alone, Outcome 4 Death within 12 months.

Techniques Used:

Comparison 4 Treatment of first rejection (B cell): rituximab + steroids versus steroids alone, Outcome 2 Additional treatment required.
Figure Legend Snippet: Comparison 4 Treatment of first rejection (B cell): rituximab + steroids versus steroids alone, Outcome 2 Additional treatment required.

Techniques Used:

Comparison 4 Treatment of first rejection (B cell): rituximab + steroids versus steroids alone, Outcome 1 Failure of reversal of acute rejection.
Figure Legend Snippet: Comparison 4 Treatment of first rejection (B cell): rituximab + steroids versus steroids alone, Outcome 1 Failure of reversal of acute rejection.

Techniques Used:

Comparison 4 Treatment of first rejection (B cell): rituximab + steroids versus steroids alone, Outcome 5 Treatment adverse events.
Figure Legend Snippet: Comparison 4 Treatment of first rejection (B cell): rituximab + steroids versus steroids alone, Outcome 5 Treatment adverse events.

Techniques Used:

17) Product Images from "Impact of Rituximab (Rituxan) on the Treatment of B-Cell Non-Hodgkin's Lymphoma"

Article Title: Impact of Rituximab (Rituxan) on the Treatment of B-Cell Non-Hodgkin's Lymphoma

Journal: Pharmacy and Therapeutics

doi:

Overall survival according to treatment regimen in diffuse large B-cell lymphoma. Results of a randomized trial of non-rituximab containing regimens as historical controls for British Columbia outcome data with R-CHOP. MACOP-B = methotrexate, Adriamycin,
Figure Legend Snippet: Overall survival according to treatment regimen in diffuse large B-cell lymphoma. Results of a randomized trial of non-rituximab containing regimens as historical controls for British Columbia outcome data with R-CHOP. MACOP-B = methotrexate, Adriamycin,

Techniques Used:

18) Product Images from "Human CD59 inhibitor sensitizes rituximab resistant lymphoma cells to complement-mediated cytolysis"

Article Title: Human CD59 inhibitor sensitizes rituximab resistant lymphoma cells to complement-mediated cytolysis

Journal: Cancer research

doi: 10.1158/0008-5472.CAN-10-3016

rILYd4 is an effective adjuvant for rituximab against RR cells or CLL cells
Figure Legend Snippet: rILYd4 is an effective adjuvant for rituximab against RR cells or CLL cells

Techniques Used:

Expression of CD20, CD46, CD59, CD55 and CD48 on CD59-enriched CS cells with and without ILY pretreatment and their response to CDC effect induced by rituximab in vitro
Figure Legend Snippet: Expression of CD20, CD46, CD59, CD55 and CD48 on CD59-enriched CS cells with and without ILY pretreatment and their response to CDC effect induced by rituximab in vitro

Techniques Used: Expressing, In Vitro

Expression of CD59 on OR and RR cells with or without ILY pretreatment and their response to rituximab-mediated CDC in vitro
Figure Legend Snippet: Expression of CD59 on OR and RR cells with or without ILY pretreatment and their response to rituximab-mediated CDC in vitro

Techniques Used: Expressing, In Vitro

19) Product Images from "A transgene-encoded cell surface polypeptide for selection, in vivo tracking, and ablation of engineered cells"

Article Title: A transgene-encoded cell surface polypeptide for selection, in vivo tracking, and ablation of engineered cells

Journal: Blood

doi: 10.1182/blood-2011-02-337360

Use of EGFRt as a target for Erbitux-mediated ADCC of engineered cells. (A) 51 Cr-labeled huEGFRt-selected T cells (inset depicts huEGFRt expression) were mixed with 1 μg/mL of Erbitux or Rituxan as a negative control before addition of GM-CSF–stimulated
Figure Legend Snippet: Use of EGFRt as a target for Erbitux-mediated ADCC of engineered cells. (A) 51 Cr-labeled huEGFRt-selected T cells (inset depicts huEGFRt expression) were mixed with 1 μg/mL of Erbitux or Rituxan as a negative control before addition of GM-CSF–stimulated

Techniques Used: Labeling, Expressing, Negative Control

20) Product Images from "High-dose methotrexate and rituximab with deferred radiotherapy for newly diagnosed primary B-cell CNS lymphoma"

Article Title: High-dose methotrexate and rituximab with deferred radiotherapy for newly diagnosed primary B-cell CNS lymphoma

Journal: Neuro-Oncology

doi: 10.1093/neuonc/noq011

Progression free and overall survival for patients with newly diagnosed primary CNS lymphoma treated with high-dose methotrexate and rituximab.
Figure Legend Snippet: Progression free and overall survival for patients with newly diagnosed primary CNS lymphoma treated with high-dose methotrexate and rituximab.

Techniques Used:

Related Articles

other:

Article Title: Polyclonal and monoclonal antibodies for treating acute rejection episodes in kidney transplant recipients
Article Snippet: Two studies looking at humoral rejection using rituximab; (20 participants) compared rituximab with steroid‐to‐steroid alone, and (38 participants) compared rituximab with placebo. used anti‐CD 20 rituximab manufactured by BIOGEN‐IDEC Pharmaceuticals and Genentech Inc.

Article Title: Responsiveness to reduced dosage of rituximab in Chinese patients with neuromyelitis optica
Article Snippet: All 5 patients were treated with rituximab (Biogen-Idec, Cambridge, MA, and Genentech, San Francisco, CA) 100 mg (equivalent of 50–59 mg/m2 ) IV, one infusion per week for 3 consecutive weeks ( ).

Concentration Assay:

Article Title: Ofatumumab Exhibits Enhanced In Vitro and In Vivo Activity Compared to Rituximab in Preclinical Models of Mantle Cell Lymphoma
Article Snippet: Flow-cytometric analysis was performed using the FCS express software version 3 for windows (De Novo Software). .. Rituximab (Biogen Idec and Genentech) was obtained from the RPCI Pharmacy Department at a stock concentration of 10 mg/mL. .. Chimeric anti-human Her-2 neu (trastuzumab) provided by Genentech was used as an isotype control.

Mouse Assay:

Article Title: Rituximab specifically depletes short-lived autoreactive plasma cells in a mouse model of inflammatory arthritis
Article Snippet: This unanticipated expression may be related to the relatively immature status of plasmablasts, or it may be a property of the hCD20 BAC transgene. .. Therefore, we assessed the display of endogenous mouse CD20 on GPI-positive plasma cells in K/BxN vs. BxN mice with a well-characterized biotinylated anti-mouse CD20 antibody (clone 18B12 IgG2b; Biogen). .. Both of the GPI-positive plasma cell populations in SP and LNs expressed mouse CD20, although the expression levels were about one-third to one-half of that on standard B cells ( ).

Isolation:

Article Title: A transgene-encoded cell surface polypeptide for selection, in vivo tracking, and ablation of engineered cells
Article Snippet: .. Antibody-dependent cell-mediated cytotoxicity (ADCC) was determined using up to 2.5 × 105 freshly isolated PBMCs that had been prestimulated overnight with 10 ng/mL huGM-CSF (R & D Systems) as effector cells in cocultures with 5 × 103 51 Cr-labeled huEGFRt+ T cells as targets, with or without 1 μg/mL of either Erbitux or rituximab (Rituxan Genentec/Biogen IDEC). ..

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    Biogen Inc chimeric mouse human anti cd20 antibody rituximab c2b8
    Characterization of purified <t>rituximab</t> minibody and scFv-Fc DM. A) Schematic presentations of the minibody and scFv-Fc DM. Both proteins assemble into covalent bound homodimers through cysteines in the hinge region. The two mutations (H310A and H435Q) present in the Fc region are indicated. V L = variable light. V H = variable heavy. C H = constant heavy. B) Coomassie stained SDS-PAGE of purified proteins under non-reducing (lanes 2) and reducing (lanes 3) conditions. Lane 1 = molecular marker. C) Size-exclusion chromatography analysis of purified minibody and scFv-Fc DM. D) Flow cytometric analysis of <t>CD20</t> binding by rituximab antibody fragments. Purified proteins were assayed for binding to 38C13-huCD20 cells. Bound protein was detected with phycoerythrin (PE) conjugated goat anti-human (Fc specific) antibodies. Rituximab was used as positive control and secondary antibody alone as negative control.
    Chimeric Mouse Human Anti Cd20 Antibody Rituximab C2b8, supplied by Biogen Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Biogen Inc rituximab
    Kinetics of the B-cell population in patients treated with repeated dosage of 100 mg <t>rituximab</t>
    Rituximab, supplied by Biogen Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Characterization of purified rituximab minibody and scFv-Fc DM. A) Schematic presentations of the minibody and scFv-Fc DM. Both proteins assemble into covalent bound homodimers through cysteines in the hinge region. The two mutations (H310A and H435Q) present in the Fc region are indicated. V L = variable light. V H = variable heavy. C H = constant heavy. B) Coomassie stained SDS-PAGE of purified proteins under non-reducing (lanes 2) and reducing (lanes 3) conditions. Lane 1 = molecular marker. C) Size-exclusion chromatography analysis of purified minibody and scFv-Fc DM. D) Flow cytometric analysis of CD20 binding by rituximab antibody fragments. Purified proteins were assayed for binding to 38C13-huCD20 cells. Bound protein was detected with phycoerythrin (PE) conjugated goat anti-human (Fc specific) antibodies. Rituximab was used as positive control and secondary antibody alone as negative control.

    Journal: Journal of nuclear medicine : official publication, Society of Nuclear Medicine

    Article Title: Recombinant anti-CD20 antibody fragments for microPET imaging of B-cell lymphoma

    doi: 10.2967/jnumed.108.060426

    Figure Lengend Snippet: Characterization of purified rituximab minibody and scFv-Fc DM. A) Schematic presentations of the minibody and scFv-Fc DM. Both proteins assemble into covalent bound homodimers through cysteines in the hinge region. The two mutations (H310A and H435Q) present in the Fc region are indicated. V L = variable light. V H = variable heavy. C H = constant heavy. B) Coomassie stained SDS-PAGE of purified proteins under non-reducing (lanes 2) and reducing (lanes 3) conditions. Lane 1 = molecular marker. C) Size-exclusion chromatography analysis of purified minibody and scFv-Fc DM. D) Flow cytometric analysis of CD20 binding by rituximab antibody fragments. Purified proteins were assayed for binding to 38C13-huCD20 cells. Bound protein was detected with phycoerythrin (PE) conjugated goat anti-human (Fc specific) antibodies. Rituximab was used as positive control and secondary antibody alone as negative control.

    Article Snippet: The chimeric (mouse/human) anti-CD20 antibody rituximab C2B8 (Rituxan® , Genentech/Biogen-IDEC, San Francisco, CA) has become a mainstay in the treatment of B cell NHL, achieving high response rates in low-grade B cell lymphomas , and improvements in survival in both low-grade and aggressive lymphomas when combined with chemotherapy ( , ).

    Techniques: Purification, Staining, SDS Page, Marker, Size-exclusion Chromatography, Flow Cytometry, Binding Assay, Positive Control, Negative Control

    Kinetics of the B-cell population in patients treated with repeated dosage of 100 mg rituximab

    Journal: Neurology

    Article Title: Responsiveness to reduced dosage of rituximab in Chinese patients with neuromyelitis optica

    doi: 10.1212/WNL.0b013e3182a1aac7

    Figure Lengend Snippet: Kinetics of the B-cell population in patients treated with repeated dosage of 100 mg rituximab

    Article Snippet: All 5 patients were treated with rituximab (Biogen-Idec, Cambridge, MA, and Genentech, San Francisco, CA) 100 mg (equivalent of 50–59 mg/m2 ) IV, one infusion per week for 3 consecutive weeks ( ).

    Techniques:

    Brain and spinal cord MRIs of a patient treated with repeated dosages of 100 mg rituximab

    Journal: Neurology

    Article Title: Responsiveness to reduced dosage of rituximab in Chinese patients with neuromyelitis optica

    doi: 10.1212/WNL.0b013e3182a1aac7

    Figure Lengend Snippet: Brain and spinal cord MRIs of a patient treated with repeated dosages of 100 mg rituximab

    Article Snippet: All 5 patients were treated with rituximab (Biogen-Idec, Cambridge, MA, and Genentech, San Francisco, CA) 100 mg (equivalent of 50–59 mg/m2 ) IV, one infusion per week for 3 consecutive weeks ( ).

    Techniques:

    Figure 1. (A) The structure and topology of CD20 and the epitopes recognized by rituximab, ofatumumab and GA101. (B) Sequence alignment of CD20 epitopes recognized by CD20 antibodies based on published information. Core epitope residues are boxed

    Journal: mAbs

    Article Title: Epitope interactions of monoclonal antibodies targeting CD20 and their relationship to functional properties

    doi: 10.4161/mabs.22771

    Figure Lengend Snippet: Figure 1. (A) The structure and topology of CD20 and the epitopes recognized by rituximab, ofatumumab and GA101. (B) Sequence alignment of CD20 epitopes recognized by CD20 antibodies based on published information. Core epitope residues are boxed

    Article Snippet: Rituximab (MabThera® ; Rituxan® , Roche/Genentech/Biogen IDEC) was the first monoclonal antibody to be approved for the treatment of lymphoma, and it has changed the treatment of non-Hodgkin lymphoma (NHL) and chronic lymphocytic leukemia (CLL), particularly in combination with chemotherapy where it has been shown to improve survival compared with chemotherapy alone.

    Techniques: Sequencing

    Figure 6. Three-dimensional models of A) rituximab and B) GA101. GA101 binds to the same binding epitope region of CD20 as rituximab, but in a different binding orientation. The molecular models were created by combining known structural data

    Journal: mAbs

    Article Title: Epitope interactions of monoclonal antibodies targeting CD20 and their relationship to functional properties

    doi: 10.4161/mabs.22771

    Figure Lengend Snippet: Figure 6. Three-dimensional models of A) rituximab and B) GA101. GA101 binds to the same binding epitope region of CD20 as rituximab, but in a different binding orientation. The molecular models were created by combining known structural data

    Article Snippet: Rituximab (MabThera® ; Rituxan® , Roche/Genentech/Biogen IDEC) was the first monoclonal antibody to be approved for the treatment of lymphoma, and it has changed the treatment of non-Hodgkin lymphoma (NHL) and chronic lymphocytic leukemia (CLL), particularly in combination with chemotherapy where it has been shown to improve survival compared with chemotherapy alone.

    Techniques: Binding Assay

    Figure 5. Comparison of A) rituximab (Type I) and B) GA101 (Type II) crystal structures in complex with CD20 peptide.29 While for rituximab N171 is deeply immersed and N176 has no contacts with the rituximab CDRs, N171 is not deeply immersed in

    Journal: mAbs

    Article Title: Epitope interactions of monoclonal antibodies targeting CD20 and their relationship to functional properties

    doi: 10.4161/mabs.22771

    Figure Lengend Snippet: Figure 5. Comparison of A) rituximab (Type I) and B) GA101 (Type II) crystal structures in complex with CD20 peptide.29 While for rituximab N171 is deeply immersed and N176 has no contacts with the rituximab CDRs, N171 is not deeply immersed in

    Article Snippet: Rituximab (MabThera® ; Rituxan® , Roche/Genentech/Biogen IDEC) was the first monoclonal antibody to be approved for the treatment of lymphoma, and it has changed the treatment of non-Hodgkin lymphoma (NHL) and chronic lymphocytic leukemia (CLL), particularly in combination with chemotherapy where it has been shown to improve survival compared with chemotherapy alone.

    Techniques:

    Figure 4. Published crystal structures of CD20 antibodies. A) rituximab-CD20 complex,48 B) ofatumumab (no co-crystal structure is available),50 C) 2H7-CD20 complex,49 and D) GA101-CD20 complex.29 The heavy chain is colored in darker shades, the

    Journal: mAbs

    Article Title: Epitope interactions of monoclonal antibodies targeting CD20 and their relationship to functional properties

    doi: 10.4161/mabs.22771

    Figure Lengend Snippet: Figure 4. Published crystal structures of CD20 antibodies. A) rituximab-CD20 complex,48 B) ofatumumab (no co-crystal structure is available),50 C) 2H7-CD20 complex,49 and D) GA101-CD20 complex.29 The heavy chain is colored in darker shades, the

    Article Snippet: Rituximab (MabThera® ; Rituxan® , Roche/Genentech/Biogen IDEC) was the first monoclonal antibody to be approved for the treatment of lymphoma, and it has changed the treatment of non-Hodgkin lymphoma (NHL) and chronic lymphocytic leukemia (CLL), particularly in combination with chemotherapy where it has been shown to improve survival compared with chemotherapy alone.

    Techniques:

    Figure 3. Hypothetical model for CD20 binding of Type I and Type II CD20 antibodies explaining the impact of FcγRIIb on internalization. A) Type I antibodies such as rituximab may bind to CD20 in a conformation that allows simultaneous

    Journal: mAbs

    Article Title: Epitope interactions of monoclonal antibodies targeting CD20 and their relationship to functional properties

    doi: 10.4161/mabs.22771

    Figure Lengend Snippet: Figure 3. Hypothetical model for CD20 binding of Type I and Type II CD20 antibodies explaining the impact of FcγRIIb on internalization. A) Type I antibodies such as rituximab may bind to CD20 in a conformation that allows simultaneous

    Article Snippet: Rituximab (MabThera® ; Rituxan® , Roche/Genentech/Biogen IDEC) was the first monoclonal antibody to be approved for the treatment of lymphoma, and it has changed the treatment of non-Hodgkin lymphoma (NHL) and chronic lymphocytic leukemia (CLL), particularly in combination with chemotherapy where it has been shown to improve survival compared with chemotherapy alone.

    Techniques: Binding Assay

    Expression of CD20, CD55, and CD59 in MCL cell lines compared with rituximab-sensitive Raji and rituximab-resistant Raji 4RH cell lines. The surface density of CD20, CD55, and CD59 was determined using Imagestream. MCL cell lines exhibit similar density of CD20 expression on their cell membrane when compared with the rituximab-sensitive Raji cell line. However, the complement inhibitory proteins CD55 and CD59 are present at increased surface density compared with Raji cells, similar to the rituximab-resistant Raji 4RH cell line.

    Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

    Article Title: Ofatumumab Exhibits Enhanced In Vitro and In Vivo Activity Compared to Rituximab in Preclinical Models of Mantle Cell Lymphoma

    doi: 10.1158/1078-0432.CCR-15-0056

    Figure Lengend Snippet: Expression of CD20, CD55, and CD59 in MCL cell lines compared with rituximab-sensitive Raji and rituximab-resistant Raji 4RH cell lines. The surface density of CD20, CD55, and CD59 was determined using Imagestream. MCL cell lines exhibit similar density of CD20 expression on their cell membrane when compared with the rituximab-sensitive Raji cell line. However, the complement inhibitory proteins CD55 and CD59 are present at increased surface density compared with Raji cells, similar to the rituximab-resistant Raji 4RH cell line.

    Article Snippet: Rituximab (Biogen Idec and Genentech) was obtained from the RPCI Pharmacy Department at a stock concentration of 10 mg/mL.

    Techniques: Expressing

    Ofatumumab and rituximab induce a similar degree of ADCC-associated cell lyis. MCL tumor cell lines were tested for ADCC cell lysis using a 51 Cr release assay using effector cells from normal healthy donors at a ratio of 40:1. In all MCL cell lines tested, ofatumumab and rituximab induced similar levels of ADCC lysis at a dose of 10 μg/mL.

    Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

    Article Title: Ofatumumab Exhibits Enhanced In Vitro and In Vivo Activity Compared to Rituximab in Preclinical Models of Mantle Cell Lymphoma

    doi: 10.1158/1078-0432.CCR-15-0056

    Figure Lengend Snippet: Ofatumumab and rituximab induce a similar degree of ADCC-associated cell lyis. MCL tumor cell lines were tested for ADCC cell lysis using a 51 Cr release assay using effector cells from normal healthy donors at a ratio of 40:1. In all MCL cell lines tested, ofatumumab and rituximab induced similar levels of ADCC lysis at a dose of 10 μg/mL.

    Article Snippet: Rituximab (Biogen Idec and Genentech) was obtained from the RPCI Pharmacy Department at a stock concentration of 10 mg/mL.

    Techniques: Lysis, Release Assay

    In MCL cells derived from patient samples, ofatumumab induced similar or more cell lysis by CDC compared with rituximab. B cells were isolated from pretreatment tumor samples ( n = 4) obtained from MCL patients and tested for monoclonal antibody response by CellTiter Glo assay using rituximab or ofatumumab at a concentration of 10 μg/mL in the presence of human serum as a source of complement. Two of the four samples exhibit a statistically significant decrease in viability following ofatumumab exposure as compared to rituximab. The remaining samples show a similar response to each antibody.

    Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

    Article Title: Ofatumumab Exhibits Enhanced In Vitro and In Vivo Activity Compared to Rituximab in Preclinical Models of Mantle Cell Lymphoma

    doi: 10.1158/1078-0432.CCR-15-0056

    Figure Lengend Snippet: In MCL cells derived from patient samples, ofatumumab induced similar or more cell lysis by CDC compared with rituximab. B cells were isolated from pretreatment tumor samples ( n = 4) obtained from MCL patients and tested for monoclonal antibody response by CellTiter Glo assay using rituximab or ofatumumab at a concentration of 10 μg/mL in the presence of human serum as a source of complement. Two of the four samples exhibit a statistically significant decrease in viability following ofatumumab exposure as compared to rituximab. The remaining samples show a similar response to each antibody.

    Article Snippet: Rituximab (Biogen Idec and Genentech) was obtained from the RPCI Pharmacy Department at a stock concentration of 10 mg/mL.

    Techniques: Derivative Assay, Lysis, Isolation, Glo Assay, Concentration Assay

    Ofatumumab and rituximab exhibit modest direct antitumor activity against MCL cell lines. MCL cell lines were utilized in alamarBlue reduction assays to investigate the direct antilymphoma activity of each antibody. Each antibody exhibited a time-dependent decrease in MCL cell viability following exposure to 10 μg/mL of each respective antibody in the presence of a cross-linking IgG antibody. With the exception of the Mino cell line at the 72-hour time point, all cell lines demonstrated similar degrees of direct antitumor activity with ofatumumab and rituximab at the dose tested (*, P

    Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

    Article Title: Ofatumumab Exhibits Enhanced In Vitro and In Vivo Activity Compared to Rituximab in Preclinical Models of Mantle Cell Lymphoma

    doi: 10.1158/1078-0432.CCR-15-0056

    Figure Lengend Snippet: Ofatumumab and rituximab exhibit modest direct antitumor activity against MCL cell lines. MCL cell lines were utilized in alamarBlue reduction assays to investigate the direct antilymphoma activity of each antibody. Each antibody exhibited a time-dependent decrease in MCL cell viability following exposure to 10 μg/mL of each respective antibody in the presence of a cross-linking IgG antibody. With the exception of the Mino cell line at the 72-hour time point, all cell lines demonstrated similar degrees of direct antitumor activity with ofatumumab and rituximab at the dose tested (*, P

    Article Snippet: Rituximab (Biogen Idec and Genentech) was obtained from the RPCI Pharmacy Department at a stock concentration of 10 mg/mL.

    Techniques: Activity Assay

    Ofatumumab induces significantly higher levels of CDC-associated cell lysis than rituximab. MCL tumor cell lines were tested for CDC cell lysis using a 51 Cr release assay with human serum from normal healthy donors as a source of complement. In all MCL cell lines tested, with the exception of Granta, ofatumumab was more effective than rituximab at inducing CDC lysis at a dose of 10 μg/mL. (*, P

    Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

    Article Title: Ofatumumab Exhibits Enhanced In Vitro and In Vivo Activity Compared to Rituximab in Preclinical Models of Mantle Cell Lymphoma

    doi: 10.1158/1078-0432.CCR-15-0056

    Figure Lengend Snippet: Ofatumumab induces significantly higher levels of CDC-associated cell lysis than rituximab. MCL tumor cell lines were tested for CDC cell lysis using a 51 Cr release assay with human serum from normal healthy donors as a source of complement. In all MCL cell lines tested, with the exception of Granta, ofatumumab was more effective than rituximab at inducing CDC lysis at a dose of 10 μg/mL. (*, P

    Article Snippet: Rituximab (Biogen Idec and Genentech) was obtained from the RPCI Pharmacy Department at a stock concentration of 10 mg/mL.

    Techniques: Lysis, Release Assay