cd79  (Novus Biologicals)

Journal: Molecular Cancer Therapeutics

Article Title: Broadening the Therapeutic Window of ADCs Using Site-Specific Bioconjugation Showcased by an MMAE-Containing Peptide Linker in a CD79b-Targeting ADC

doi: 10.1158/1535-7163.MCT-24-0983

Figure Lengend Snippet: Concept of novel one-step and site-specific MTG technology and manufacturing of anti–CD79b-RKAA-MMAE. A, Schematic drawing of the novel MTG-based conjugation technology using a linker-payload composed of a small RKAA peptide linker and the cytotoxic payload MMAE. Functionalization occurs at the Q295 residue of the native, glycosylated polatuzumab antibody. RKAA-MMAE is attached in a single step using wild-type MTG. B, Linker-payload structure comprised the RKAA-peptide linker with an acetylated N-terminus, a PABC self-immolative spacer, and MMAE. C, Schematic comparison of anti–CD79b-RKAA-MMAE generated by one-step site-specific MTG conjugation and FDA-approved PV.

Article Snippet: Plasma concentrations were determined by ELISA using His-tagged human CD79b (Sino Biological) as the capturing agent, coated on Ni-NTA HisSorb plates (Qiagen).

Techniques: Conjugation Assay, Residue, Comparison, Generated

Journal: Molecular Cancer Therapeutics

Article Title: Broadening the Therapeutic Window of ADCs Using Site-Specific Bioconjugation Showcased by an MMAE-Containing Peptide Linker in a CD79b-Targeting ADC

doi: 10.1158/1535-7163.MCT-24-0983

Figure Lengend Snippet: Analytic characterization of anti–CD79b-RKAA-MMAE. A, LC/MS spectra of the heavy chain (HC) and light chain (LC) of glycosylated anti–CD79b-RKAA-MMAE under reducing conditions. Main peaks showing single conjugation of HC (left graph) and unconjugated LC (right graph). B, Intact LC/MS. Main peaks showing DAR 2 ADC. C, Reverse-phase UHPLC analysis of reduced anti–CD79b-RKAA-MMAE. D, Analysis of monomer content by SEC. E, Comparison of HIC profiles of unconjugated polatuzumab antibody, anti–CD79b-RKAA-MMAE, and PV. Different DAR species of the preparations are indicated.

Article Snippet: Plasma concentrations were determined by ELISA using His-tagged human CD79b (Sino Biological) as the capturing agent, coated on Ni-NTA HisSorb plates (Qiagen).

Techniques: Liquid Chromatography with Mass Spectroscopy, Conjugation Assay, Comparison

Journal: Molecular Cancer Therapeutics

Article Title: Broadening the Therapeutic Window of ADCs Using Site-Specific Bioconjugation Showcased by an MMAE-Containing Peptide Linker in a CD79b-Targeting ADC

doi: 10.1158/1535-7163.MCT-24-0983

Figure Lengend Snippet: In vitro evaluation of binding, toxicity, and stability of anti–CD79b-RKAA-MMAE. A, Binding comparison of unconjugated polatuzumab, anti–CD79b-RKAA-MMAE, and PV on recombinant CD79b by ELISA. The graph shows mean ± SD of technical duplicates. B, Representative assay to evaluate target binding on CD79b-positive BJAB cells. C, Evaluation of Fc functionality by ELISA on immobilized FcγRI (CD64). D, In vitro cytotoxicity assay on CD79b-expressing Granta-519 cells. Comparison of anti–CD79b-RKAA-MMAE (DAR 2) and PV (DAR 3.5) at equal ADC concentrations. E, Toxicity on CD79b-positive WSU-DLCL2 tumor cells. F, In vitro cytotoxicity assay on target-negative HT cells. G, All cytotoxicity graphs show mean ± SD of technical duplicates. A representative serum stability assay of anti–CD79b-RKAA-MMAE in human, cynomolgus monkey, and mouse IgG-depleted sera at 37°C for 7 days. DAR was assessed by LC/MS at different time points. H, Evaluation of PV stability in human, cynomolgus monkey, and mouse IgG-depleted sera. MFI, mean fluorescence intensity; OD, optical density.

Article Snippet: Plasma concentrations were determined by ELISA using His-tagged human CD79b (Sino Biological) as the capturing agent, coated on Ni-NTA HisSorb plates (Qiagen).

Techniques: In Vitro, Binding Assay, Comparison, Recombinant, Enzyme-linked Immunosorbent Assay, Cytotoxicity Assay, Expressing, Stability Assay, Liquid Chromatography with Mass Spectroscopy, Fluorescence

Journal: Molecular Cancer Therapeutics

Article Title: Broadening the Therapeutic Window of ADCs Using Site-Specific Bioconjugation Showcased by an MMAE-Containing Peptide Linker in a CD79b-Targeting ADC

doi: 10.1158/1535-7163.MCT-24-0983

Figure Lengend Snippet: Enzymatic cleavage of anti–CD79b-RKAA-MMAE. Comparison of the cleavage rate of anti–CD79b-RKAA-MMAE and PV using human cathepsin B ( A ), mouse Ces1c ( B ), or HLL extract ( C ). D, Schematic overview of the cleavage mechanism of anti–CD79b-RKAA-MMAE by proteases and subsequent payload release and liberation of free MMAE by 1,6-elimination of PABC.

Article Snippet: Plasma concentrations were determined by ELISA using His-tagged human CD79b (Sino Biological) as the capturing agent, coated on Ni-NTA HisSorb plates (Qiagen).

Techniques: Comparison

Journal: Molecular Cancer Therapeutics

Article Title: Broadening the Therapeutic Window of ADCs Using Site-Specific Bioconjugation Showcased by an MMAE-Containing Peptide Linker in a CD79b-Targeting ADC

doi: 10.1158/1535-7163.MCT-24-0983

Figure Lengend Snippet: Pharmacokinetic studies in Swiss wild-type mice and Sprague Dawley rats. A, Healthy female Swiss wild-type mice were injected intravenously with 5 mg/kg unconjugated polatuzumab, anti–CD79b-RKAA-MMAE, or PV ( n = 5 per group). Plasma was collected at different time points after injection, and total IgG and intact ADC concentrations were determined by ELISA. Data represent mean plasma concentrations (±SD). B, Male Sprague Dawley rats were injected intravenously with 10 mg/kg of anti–CD79b-RKAA-MMAE (top graph) or 10 mg/kg of PV (bottom graph) in a repeat-dose study ( n = 5 per group). Animals were injected weekly for a total of four injections. Graphs show plasma concentration–time curves after the fourth dose, including the predose concentration measurement (0 hours).

Article Snippet: Plasma concentrations were determined by ELISA using His-tagged human CD79b (Sino Biological) as the capturing agent, coated on Ni-NTA HisSorb plates (Qiagen).

Techniques: Injection, Clinical Proteomics, Enzyme-linked Immunosorbent Assay, Concentration Assay

Journal: Molecular Cancer Therapeutics

Article Title: Broadening the Therapeutic Window of ADCs Using Site-Specific Bioconjugation Showcased by an MMAE-Containing Peptide Linker in a CD79b-Targeting ADC

doi: 10.1158/1535-7163.MCT-24-0983

Figure Lengend Snippet: Therapeutic efficacy of anti–CD79b-RKAA-MMAE in different xenograft models. Treatment was started when tumors reached a size of approximately 200 mm 3 . Mice were allocated into different groups using a nonrandom stratification protocol. Data represent mean tumor volumes (±SEM). A, CB17-SCID mice were challenged with 20 × 10 6 Granta-519 tumor cells and received a single injection (black arrow) of 2.1 mg/kg anti–CD79b-RKAA-MMAE (corresponding to 20 μg/kg of payload), 2.1 mg/kg PV (corresponding to 40 μg/kg of payload), or PBS, intravenously into the lateral tail vein ( n = 8 per group). B, Treatment of the Granta-519 xenograft model at equal payload doses of 10 µg/kg, corresponding to 1 mg/kg of anti–CD79b-RKAA-MMAE or 0.53 mg/kg of PV ( n = 8 per group). C, Administration of 1.5 mg/kg (15 μg/kg of payload) of anti–CD79b-RKAA-MMAE or nonbinding control IgG1-RKAA-MMAE to Granta-519–bearing CB17-SCID mice. D, CB17-SCID mice were challenged with 20 × 10 6 Ramos Burkitt lymphoma cells. Anti–CD79b-RKAA-MMAE (2.5 mg/kg) or PV (1.43 mg/kg; both corresponding to 25 μg/kg of payload) was used for treatment ( n = 6 per group). PBS was used as a negative control. *, P < 0.05; ****, P < 0.0001. CR, complete response.

Article Snippet: Plasma concentrations were determined by ELISA using His-tagged human CD79b (Sino Biological) as the capturing agent, coated on Ni-NTA HisSorb plates (Qiagen).

Techniques: Drug discovery, Injection, Control, Negative Control

Journal: Animals : an Open Access Journal from MDPI

Article Title: Postpartum Uterine Involution in Cows: Quantitative Assessment of Structural Remodeling and Immune Cell Infiltration

doi: 10.3390/ani15172520

Figure Lengend Snippet: Bland–Altman plot showing the differences between manual and automated quantification of positively marked cells in the bovine endometrium for CD3 ( A ), CD79 ( B ), CD204 ( C ) and CD335 ( D ) during the postpartum period, plotted against the average of the two methods (pink dots). The bias is indicated by the distance between the zero-difference line ( X -axis) and the solid green line. The limits of agreement are represented by the dashed lines parallel to the X -axis. Numerical values for the bias and limits of agreement are displayed adjacent to their respective lines.

Article Snippet: CD79 , Novus Biologicals; Centennial, CO, USA (NB-1006434 7) , MoM , 1:100.

Techniques:

Journal: Animals : an Open Access Journal from MDPI

Article Title: Postpartum Uterine Involution in Cows: Quantitative Assessment of Structural Remodeling and Immune Cell Infiltration

doi: 10.3390/ani15172520

Figure Lengend Snippet: Endometrial biopsies of bovine uterus. Immunostaining and quantification of T cells using CD3 (1:100) and B-cells using CD79 (1:100) and counterstained with Hematoxylin. ( A – D ) Negative controls, stained with Hematoxylin only, are the same for all antibodies and were used to confirm tissue morphology preservation. ( E – H ) Endometrial biopsies of bovine uterus. Immunostaining for CD3 on Days 2, 7, 21, and 35 postpartum, respectively. ( I – L ) Endometrial biopsies of bovine uterus. Immunostaining for CD79 on Days 2, 7, 21, and 35 postpartum, respectively. The images were obtained using Aperio ImageScope x64 12.4.6 software (Leica Biosystems), magnification 400x, scale 25 µm. ( M , N ) Manual ( M ) and automatic ( N ) counting of CD3-positive cells. ( O , P ) Manual ( O ) and automatic ( P ) counting of CD79-positive cells. Different letters are statistically significant ( p < 0.05). Results are presented as mean ± standard error of the mean (S.E.M.).

Article Snippet: CD79 , Novus Biologicals; Centennial, CO, USA (NB-1006434 7) , MoM , 1:100.

Techniques: Immunostaining, Staining, Preserving, Software

Journal: iScience

Article Title: PI3K-dependent reprogramming of hexokinase isoforms controls glucose metabolism and functional responses of B lymphocytes

doi: 10.1016/j.isci.2024.110939

Figure Lengend Snippet:

Article Snippet: MB1-Cre (B6.C(Cg)-CD79 tm1(cre)Reth /Ehob/J , Jackson Laboratory , 020505.

Techniques: Activation Assay, Marker, Recombinant, Staining, Cell Isolation, Software

Journal: Breast Cancer Research : BCR

Article Title: Factors associated with engraftment success of patient-derived xenografts of breast cancer

doi: 10.1186/s13058-024-01794-w

Figure Lengend Snippet: Artificial intelligence-assesed classification of patches. A . Adipose; B . Background; C . Necrosis; D . Ductal carcinoma in situ, classified as carcinoma; E . Lobular carcinoma in situ, classified as carcinoma; F . Invasive ductal carcinoma, classified as carcinoma; G . Invasive lobular carcinoma, classified as carcinoma; H . Stroma; I. Terminal ductal lobular unit; J. Tumor-infiltrating lymphocytes trained with a segmentation model. Left, hematoxylin and eosin (H&E) stained slide, X100, original magnification. Middle, Cocktail immunohistochemistry (IHC) for identification of lymphocytes, X100, CD3, CD20, and CD79 cocktail IHC, original magnification. Right, red annotation indicating the area of cocktail IHC-stained lymphocytes in the H&E slides (annotated digitally processed image, original magnification, X100). K-L . Representative H&E from successful ( K ) and failed ( L ) PDX graft cases and their corresponding AI-model applied images. K . Abundant intratumoral necrosis and carcinoma proportions in the H&E are also highlighted in sky-blue and green in the AI-model applied image, respectively (right); L. Abundant intratumoral TILs, stroma, and TDLU identified in the H&E slide (left) are also highlighted in the AI-categorized image (right). M . algorithm applied to the carcinoma (left side) generated a tumor boundary (right side, yellow) encircling the carcinoma component (right side, green)

Article Snippet: A representative H&E slide was scanned (Fig. J, left side), de-stained, and re-stained with a cocktail of immune cell markers (Fig. J, middle): CD3 (1:50, Novocastra Laboratories, Leica Biosystems, Nussloch, Germany), CD20 (1:500, Novocastra Laboratories), and CD79 (1:200, Dako, Agilent Technologies, Santa Clara, CA, USA), using a Ventana ES automated immunohistochemistry (IHC) stainer according to the manufacturer’s protocols (Ventana ES automated IHC stainer, Tucson, AZ, USA).

Techniques: In Situ, Staining, Immunohistochemistry, Generated