mngf  (Alomone Labs)


Bioz Verified Symbol Alomone Labs is a verified supplier
Bioz Manufacturer Symbol Alomone Labs manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94

    Structured Review

    Alomone Labs mngf
    Mngf, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mngf/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mngf - by Bioz Stars, 2023-01
    94/100 stars

    Images

    mngf  (Alomone Labs)


    Bioz Verified Symbol Alomone Labs is a verified supplier
    Bioz Manufacturer Symbol Alomone Labs manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94

    Structured Review

    Alomone Labs mngf
    Mngf, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mngf/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mngf - by Bioz Stars, 2023-01
    94/100 stars

    Images

    mngf  (Alomone Labs)


    Bioz Verified Symbol Alomone Labs is a verified supplier
    Bioz Manufacturer Symbol Alomone Labs manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94

    Structured Review

    Alomone Labs mngf
    Mngf, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mngf/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mngf - by Bioz Stars, 2023-01
    94/100 stars

    Images

    mngf  (Alomone Labs)


    Bioz Verified Symbol Alomone Labs is a verified supplier
    Bioz Manufacturer Symbol Alomone Labs manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94

    Structured Review

    Alomone Labs mngf
    Mngf, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mngf/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mngf - by Bioz Stars, 2023-01
    94/100 stars

    Images

    mngf  (Alomone Labs)


    Bioz Verified Symbol Alomone Labs is a verified supplier
    Bioz Manufacturer Symbol Alomone Labs manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 86

    Structured Review

    Alomone Labs mngf
    Mature and immature NGF forms in patients with JIA and RA. (A,B) Only proNGF forms of different molecular weights but not mature NGF were detected by western blot in synovial fluids of 12 patients with JIA (40 μg total protein). (B) Blocking the anti-NGF antibody by adding mature NGF results in the disappearance of the specific proNGF bands observed in synovial fluids of three patients with JIA, as well as of the band of the 25 kDa commercial proNGF that was added as positive control. (C) In JIA and RA synovial fluids, proNGF and mature NGF <t>(mNGF)</t> concentrations were assessed using a newly developed ELISA showing that proNGF is 4.8-fold higher in JIA (n=27) and 16.8-fold higher in patients with RA (n=5) than mNGF. (D) Real-time PCR shows a very low expression of NGF mRNA <t>in</t> <t>mononuclear</t> cells obtained from peripheral blood of healthy donors (CTRL PBMC) or in mononuclear cells from peripheral blood (JIA PBMC) and from synovial fluids of patients with JIA (JIA SFMC). High NGF mRNA expression levels characterised synovial tissues (RA synovia; n=5). Fibroblast-like synoviocytes (FLS; n=3) of patients with RA express more NGF mRNA than FLS from osteoarthritis patients (OA FLS) (n=4) and control fibroblasts (CTRL FB; n=4). Results were expressed as arbitrary units (AU) and obtained after normalisation with the housekeeping gene glyceraldehyde 3-phosphate dehydrogenase (GAPDH). (E) Both unstimulated (US) and LPS-stimulated JIA SFMC do not release proNGF or mNGF in the supernatants. Commercial proNGF and mNGF were added as positive controls. (F) Western blot shows the stability of exogenous added mNGF and proNGF. After its addition to culture media, proNGF is still detected after 3 hours and undergoes a minor maturation into mNGF in the first 2 hours of incubation (left side of F). mNGF is not degraded in the first 3 hours after its addition (right side of F). Commercial proNGF and mNGF were included as positive controls. (G) Neither proNGF nor mNGF were released in culture media of US or LPS-stimulated JIA SFMC after 18 hours of incubation. Exogenous proNGF or mNGF were detected only at time of supplementation (in agreement with F) but were no longer detectable after 18 hours of incubation. Commercial proNGF and mNGF were included as positive controls. (H) ELISA instead shows that conditioned media of RA FLS (n=5) have higher concentrations of proNGF than control fibroblasts (CTRL FB; n=5) and that the concentration of proNGF is higher than mNGF after 18 hours of incubation. *p<0.05, **p<0.01, ***p<0.001. JIA, juvenile idiopathic arthritis; LPS, Lipopolysaccharide ; NGF, nerve growth factor; PBMC, mononuclear cells isolated from peripheral blood; RA rheumatoid arthritis; SFMC, mononuclear cells isolated from synovial fluids.
    Mngf, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mngf/product/Alomone Labs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mngf - by Bioz Stars, 2023-01
    86/100 stars

    Images

    1) Product Images from "ProNGF-p75NTR axis plays a proinflammatory role in inflamed joints: a novel pathogenic mechanism in chronic arthritis"

    Article Title: ProNGF-p75NTR axis plays a proinflammatory role in inflamed joints: a novel pathogenic mechanism in chronic arthritis

    Journal: RMD Open

    doi: 10.1136/rmdopen-2017-000441

    Mature and immature NGF forms in patients with JIA and RA. (A,B) Only proNGF forms of different molecular weights but not mature NGF were detected by western blot in synovial fluids of 12 patients with JIA (40 μg total protein). (B) Blocking the anti-NGF antibody by adding mature NGF results in the disappearance of the specific proNGF bands observed in synovial fluids of three patients with JIA, as well as of the band of the 25 kDa commercial proNGF that was added as positive control. (C) In JIA and RA synovial fluids, proNGF and mature NGF (mNGF) concentrations were assessed using a newly developed ELISA showing that proNGF is 4.8-fold higher in JIA (n=27) and 16.8-fold higher in patients with RA (n=5) than mNGF. (D) Real-time PCR shows a very low expression of NGF mRNA in mononuclear cells obtained from peripheral blood of healthy donors (CTRL PBMC) or in mononuclear cells from peripheral blood (JIA PBMC) and from synovial fluids of patients with JIA (JIA SFMC). High NGF mRNA expression levels characterised synovial tissues (RA synovia; n=5). Fibroblast-like synoviocytes (FLS; n=3) of patients with RA express more NGF mRNA than FLS from osteoarthritis patients (OA FLS) (n=4) and control fibroblasts (CTRL FB; n=4). Results were expressed as arbitrary units (AU) and obtained after normalisation with the housekeeping gene glyceraldehyde 3-phosphate dehydrogenase (GAPDH). (E) Both unstimulated (US) and LPS-stimulated JIA SFMC do not release proNGF or mNGF in the supernatants. Commercial proNGF and mNGF were added as positive controls. (F) Western blot shows the stability of exogenous added mNGF and proNGF. After its addition to culture media, proNGF is still detected after 3 hours and undergoes a minor maturation into mNGF in the first 2 hours of incubation (left side of F). mNGF is not degraded in the first 3 hours after its addition (right side of F). Commercial proNGF and mNGF were included as positive controls. (G) Neither proNGF nor mNGF were released in culture media of US or LPS-stimulated JIA SFMC after 18 hours of incubation. Exogenous proNGF or mNGF were detected only at time of supplementation (in agreement with F) but were no longer detectable after 18 hours of incubation. Commercial proNGF and mNGF were included as positive controls. (H) ELISA instead shows that conditioned media of RA FLS (n=5) have higher concentrations of proNGF than control fibroblasts (CTRL FB; n=5) and that the concentration of proNGF is higher than mNGF after 18 hours of incubation. *p<0.05, **p<0.01, ***p<0.001. JIA, juvenile idiopathic arthritis; LPS, Lipopolysaccharide ; NGF, nerve growth factor; PBMC, mononuclear cells isolated from peripheral blood; RA rheumatoid arthritis; SFMC, mononuclear cells isolated from synovial fluids.
    Figure Legend Snippet: Mature and immature NGF forms in patients with JIA and RA. (A,B) Only proNGF forms of different molecular weights but not mature NGF were detected by western blot in synovial fluids of 12 patients with JIA (40 μg total protein). (B) Blocking the anti-NGF antibody by adding mature NGF results in the disappearance of the specific proNGF bands observed in synovial fluids of three patients with JIA, as well as of the band of the 25 kDa commercial proNGF that was added as positive control. (C) In JIA and RA synovial fluids, proNGF and mature NGF (mNGF) concentrations were assessed using a newly developed ELISA showing that proNGF is 4.8-fold higher in JIA (n=27) and 16.8-fold higher in patients with RA (n=5) than mNGF. (D) Real-time PCR shows a very low expression of NGF mRNA in mononuclear cells obtained from peripheral blood of healthy donors (CTRL PBMC) or in mononuclear cells from peripheral blood (JIA PBMC) and from synovial fluids of patients with JIA (JIA SFMC). High NGF mRNA expression levels characterised synovial tissues (RA synovia; n=5). Fibroblast-like synoviocytes (FLS; n=3) of patients with RA express more NGF mRNA than FLS from osteoarthritis patients (OA FLS) (n=4) and control fibroblasts (CTRL FB; n=4). Results were expressed as arbitrary units (AU) and obtained after normalisation with the housekeeping gene glyceraldehyde 3-phosphate dehydrogenase (GAPDH). (E) Both unstimulated (US) and LPS-stimulated JIA SFMC do not release proNGF or mNGF in the supernatants. Commercial proNGF and mNGF were added as positive controls. (F) Western blot shows the stability of exogenous added mNGF and proNGF. After its addition to culture media, proNGF is still detected after 3 hours and undergoes a minor maturation into mNGF in the first 2 hours of incubation (left side of F). mNGF is not degraded in the first 3 hours after its addition (right side of F). Commercial proNGF and mNGF were included as positive controls. (G) Neither proNGF nor mNGF were released in culture media of US or LPS-stimulated JIA SFMC after 18 hours of incubation. Exogenous proNGF or mNGF were detected only at time of supplementation (in agreement with F) but were no longer detectable after 18 hours of incubation. Commercial proNGF and mNGF were included as positive controls. (H) ELISA instead shows that conditioned media of RA FLS (n=5) have higher concentrations of proNGF than control fibroblasts (CTRL FB; n=5) and that the concentration of proNGF is higher than mNGF after 18 hours of incubation. *p<0.05, **p<0.01, ***p<0.001. JIA, juvenile idiopathic arthritis; LPS, Lipopolysaccharide ; NGF, nerve growth factor; PBMC, mononuclear cells isolated from peripheral blood; RA rheumatoid arthritis; SFMC, mononuclear cells isolated from synovial fluids.

    Techniques Used: Western Blot, Blocking Assay, Positive Control, Enzyme-linked Immunosorbent Assay, Real-time Polymerase Chain Reaction, Expressing, Incubation, Concentration Assay, Isolation

    Ex vivo effects of proNGF and NGF on JIA mononuclear cells. (A) No changes in cell viability were observed when mononuclear cells from synovial fluid (SFMC) of patients with JIA were treated for 24 hours with proNGF or mNGF, with or without LPS stimulation. Apoptosis was assessed by Annexin V flow cytometry analysis. The results represent one of three independent experiments performed in duplicate. (B) While proNGF addition alone did not induce the production of interleukin (IL)-6 in unstimulated SFMC, in LPS-activated cells proNGF induced IL-6 release in a dose-dependent manner with a maximal effect at 200 ng/mL (n=4). (C) proNGF stimulation significantly increased IL-6, IL-1β, IL-8 and tumor necrosis factor-α (TNF-α) mRNA expression levels in LPS-stimulated SFMC (n=18) of patients with JIA after 3 hours of incubation, but did not modify IL-10 mRNA expression in either CTRL or JIA mononuclear cells. Mature NGF treatment did not significantly modify proinflammatory cytokine levels or IL-10 mRNA expression in JIA mononuclear cells, while it increased IL-10 mRNA levels in CTRL PBMC. Results are expressed as arbitrary units (AU) and obtained after normalisation with the housekeeping gene GAPDH. (D) proNGF treatment significantly increased IL-6 release after 18 hours (measured by ELISA) in LPS-stimulated SFMC from patients with JIA (n=16) compared with PBMC of healthy CTRL (n=6). Mature NGF treatment did not affect IL-6 protein levels. *p<0.05. JIA, juvenile idiopathic arthritis; LPS, Lipopolysaccharide ; NGF, nerve growth factor; PBMC, mononuclear cells isolated from peripheral blood.
    Figure Legend Snippet: Ex vivo effects of proNGF and NGF on JIA mononuclear cells. (A) No changes in cell viability were observed when mononuclear cells from synovial fluid (SFMC) of patients with JIA were treated for 24 hours with proNGF or mNGF, with or without LPS stimulation. Apoptosis was assessed by Annexin V flow cytometry analysis. The results represent one of three independent experiments performed in duplicate. (B) While proNGF addition alone did not induce the production of interleukin (IL)-6 in unstimulated SFMC, in LPS-activated cells proNGF induced IL-6 release in a dose-dependent manner with a maximal effect at 200 ng/mL (n=4). (C) proNGF stimulation significantly increased IL-6, IL-1β, IL-8 and tumor necrosis factor-α (TNF-α) mRNA expression levels in LPS-stimulated SFMC (n=18) of patients with JIA after 3 hours of incubation, but did not modify IL-10 mRNA expression in either CTRL or JIA mononuclear cells. Mature NGF treatment did not significantly modify proinflammatory cytokine levels or IL-10 mRNA expression in JIA mononuclear cells, while it increased IL-10 mRNA levels in CTRL PBMC. Results are expressed as arbitrary units (AU) and obtained after normalisation with the housekeeping gene GAPDH. (D) proNGF treatment significantly increased IL-6 release after 18 hours (measured by ELISA) in LPS-stimulated SFMC from patients with JIA (n=16) compared with PBMC of healthy CTRL (n=6). Mature NGF treatment did not affect IL-6 protein levels. *p<0.05. JIA, juvenile idiopathic arthritis; LPS, Lipopolysaccharide ; NGF, nerve growth factor; PBMC, mononuclear cells isolated from peripheral blood.

    Techniques Used: Ex Vivo, Flow Cytometry, Expressing, Incubation, Enzyme-linked Immunosorbent Assay, Isolation

    proNGF and NGF activated different intracellular pathways. (A) The neutralisation of p75NTR using either a specific anti-p75NTR antibody (a-p75; 2.5 µg/mL) or using the specific pharmacological inhibitor LM11A.31 (10 nM), decreased interleukin (IL)-6 release in LPS-activated SFMC of patients with JIA (n=16) treated with proNGF. On the contrary, the blocking of TrkA with a neutralising antibody (a-TrkA; 3 µg/mL) did not affect IL-6 release. (B) Western blot shows an increased phosphorylation of p38 and JNK after 5 minutes of proNGF stimulation in LPS-treated SFMC. mNGF treatment did not affect the phosphorylation of these downstream molecules. The result is representative of one out of three independent experiments. Results of the densitometric analysis of all experiments are expressed as arbitrary units (AU). (C) In LPS-activated SFMC of three different patients with JIA, preincubation with the inhibitor LM11A-31 (10 nM), which blocks the binding of proNGF to p75NTR, abolished the phosphorylation of p38 and JNK induced after 5 minutes of proNGF addition. Results of the densitometric analysis of all experiments are expressed as arbitrary units (AU). *p<0.05, **p<0.01, ***p<0.001. JIA, juvenile idiopathic arthritis; LPS, Lipopolysaccharide ; NGF, nerve growth factor; SFMC, mononuclear cells isolated from synovial fluids.
    Figure Legend Snippet: proNGF and NGF activated different intracellular pathways. (A) The neutralisation of p75NTR using either a specific anti-p75NTR antibody (a-p75; 2.5 µg/mL) or using the specific pharmacological inhibitor LM11A.31 (10 nM), decreased interleukin (IL)-6 release in LPS-activated SFMC of patients with JIA (n=16) treated with proNGF. On the contrary, the blocking of TrkA with a neutralising antibody (a-TrkA; 3 µg/mL) did not affect IL-6 release. (B) Western blot shows an increased phosphorylation of p38 and JNK after 5 minutes of proNGF stimulation in LPS-treated SFMC. mNGF treatment did not affect the phosphorylation of these downstream molecules. The result is representative of one out of three independent experiments. Results of the densitometric analysis of all experiments are expressed as arbitrary units (AU). (C) In LPS-activated SFMC of three different patients with JIA, preincubation with the inhibitor LM11A-31 (10 nM), which blocks the binding of proNGF to p75NTR, abolished the phosphorylation of p38 and JNK induced after 5 minutes of proNGF addition. Results of the densitometric analysis of all experiments are expressed as arbitrary units (AU). *p<0.05, **p<0.01, ***p<0.001. JIA, juvenile idiopathic arthritis; LPS, Lipopolysaccharide ; NGF, nerve growth factor; SFMC, mononuclear cells isolated from synovial fluids.

    Techniques Used: Blocking Assay, Western Blot, Binding Assay, Isolation

    mngf  (Alomone Labs)


    Bioz Verified Symbol Alomone Labs is a verified supplier
    Bioz Manufacturer Symbol Alomone Labs manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94

    Structured Review

    Alomone Labs mngf
    Mngf, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mngf/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mngf - by Bioz Stars, 2023-01
    94/100 stars

    Images

    mngf  (Alomone Labs)


    Bioz Verified Symbol Alomone Labs is a verified supplier
    Bioz Manufacturer Symbol Alomone Labs manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94

    Structured Review

    Alomone Labs mngf
    Mngf, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mngf/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mngf - by Bioz Stars, 2023-01
    94/100 stars

    Images

    mngf  (Alomone Labs)


    Bioz Verified Symbol Alomone Labs is a verified supplier
    Bioz Manufacturer Symbol Alomone Labs manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94

    Structured Review

    Alomone Labs mngf
    (a) Sequence alignment of <t>mNGF,</t> hNGF and rat NGF (Loop I and Loop II are colored in magenta and green, respectively). (b) Overall structure of NGF and structural comparisons , of Loop I (in magenta) and Loop II (in green) from mNGF (PDB_ID: 1BTG, cyan) and hNGF (PDB_ID: 1WWW, green) . This Figure was prepared with PyMOL . (c) ELISA assay with mNGF and hNGF coating (5 µg/ml) and serial dilutions of <t>parental</t> <t>mAb</t> αD11. The experiments were done in duplicate.
    Mngf, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mngf/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mngf - by Bioz Stars, 2023-01
    94/100 stars

    Images

    1) Product Images from "Single Cycle Structure-Based Humanization of an Anti-Nerve Growth Factor Therapeutic Antibody"

    Article Title: Single Cycle Structure-Based Humanization of an Anti-Nerve Growth Factor Therapeutic Antibody

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0032212

    (a) Sequence alignment of mNGF, hNGF and rat NGF (Loop I and Loop II are colored in magenta and green, respectively). (b) Overall structure of NGF and structural comparisons , of Loop I (in magenta) and Loop II (in green) from mNGF (PDB_ID: 1BTG, cyan) and hNGF (PDB_ID: 1WWW, green) . This Figure was prepared with PyMOL . (c) ELISA assay with mNGF and hNGF coating (5 µg/ml) and serial dilutions of parental mAb αD11. The experiments were done in duplicate.
    Figure Legend Snippet: (a) Sequence alignment of mNGF, hNGF and rat NGF (Loop I and Loop II are colored in magenta and green, respectively). (b) Overall structure of NGF and structural comparisons , of Loop I (in magenta) and Loop II (in green) from mNGF (PDB_ID: 1BTG, cyan) and hNGF (PDB_ID: 1WWW, green) . This Figure was prepared with PyMOL . (c) ELISA assay with mNGF and hNGF coating (5 µg/ml) and serial dilutions of parental mAb αD11. The experiments were done in duplicate.

    Techniques Used: Sequencing, Enzyme-linked Immunosorbent Assay

    (a) ELISA assay with mNGF coating (5 µg/ml); serial dilutions of parental mAb αD11, chimeric IgG1 αD11 and IgG1 hum-αD11, Protein A-Sepharose purified from transiently cotransfected CHO cells supernatants. (b) Binding curves of a range of concentrations (0.29–37.5 nM) of hNGF to immobilized parental mAb αD11 (immobilization level 1280.0 RU). (c) Binding curves of a range of concentrations (0.39–25.0 nM) of Fab hum-αD11 to immobilized hNGF (immobilization level 100.3 RU).
    Figure Legend Snippet: (a) ELISA assay with mNGF coating (5 µg/ml); serial dilutions of parental mAb αD11, chimeric IgG1 αD11 and IgG1 hum-αD11, Protein A-Sepharose purified from transiently cotransfected CHO cells supernatants. (b) Binding curves of a range of concentrations (0.29–37.5 nM) of hNGF to immobilized parental mAb αD11 (immobilization level 1280.0 RU). (c) Binding curves of a range of concentrations (0.39–25.0 nM) of Fab hum-αD11 to immobilized hNGF (immobilization level 100.3 RU).

    Techniques Used: Enzyme-linked Immunosorbent Assay, Purification, Binding Assay

    (a–d) mNGF induced differentiation of PC-12 cells. Neurite outgrowth inhibition assay: photomicrographs of PC-12 cells, treated with (a) mNGF 100 ng/ml alone or preincubated (b) with mAb αD11 (5 µg/ml) or (c) with IgG1 hum-αD11 (5 µg/ml), concentrated from stable cotransfected CHO cells supernatants. (d) Negative control: untreated PC-12 cells. (e) TrkA phosphorylation inhibition assay in 3T3 TrkA cells. 3T3 TrkA cells were incubated with the indicated combinations of mNGF, parental mAb αD11, IgG1 hum-αD11 (purified by Protein A-Sepharose) and the irrelevant mAb SV5 as a negative control. Cell lysates were separated on a 10% SDS gel and phosphorylated TrkA detected using anti-phospho-Y490 TrkA Ab. The ubiquitous band of tubulin served as gel loading control.
    Figure Legend Snippet: (a–d) mNGF induced differentiation of PC-12 cells. Neurite outgrowth inhibition assay: photomicrographs of PC-12 cells, treated with (a) mNGF 100 ng/ml alone or preincubated (b) with mAb αD11 (5 µg/ml) or (c) with IgG1 hum-αD11 (5 µg/ml), concentrated from stable cotransfected CHO cells supernatants. (d) Negative control: untreated PC-12 cells. (e) TrkA phosphorylation inhibition assay in 3T3 TrkA cells. 3T3 TrkA cells were incubated with the indicated combinations of mNGF, parental mAb αD11, IgG1 hum-αD11 (purified by Protein A-Sepharose) and the irrelevant mAb SV5 as a negative control. Cell lysates were separated on a 10% SDS gel and phosphorylated TrkA detected using anti-phospho-Y490 TrkA Ab. The ubiquitous band of tubulin served as gel loading control.

    Techniques Used: Inhibition, Negative Control, Incubation, Purification, SDS-Gel

    igg1 hum αd11 binding towards mngf  (Alomone Labs)


    Bioz Verified Symbol Alomone Labs is a verified supplier
    Bioz Manufacturer Symbol Alomone Labs manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94

    Structured Review

    Alomone Labs igg1 hum αd11 binding towards mngf
    (a) Sequence alignment of <t>mNGF,</t> hNGF and rat NGF (Loop I and Loop II are colored in magenta and green, respectively). (b) Overall structure of NGF and structural comparisons , of Loop I (in magenta) and Loop II (in green) from mNGF (PDB_ID: 1BTG, cyan) and hNGF (PDB_ID: 1WWW, green) . This Figure was prepared with PyMOL . (c) ELISA assay with mNGF and hNGF coating (5 µg/ml) and serial dilutions of parental mAb <t>αD11.</t> The experiments were done in duplicate.
    Igg1 Hum αd11 Binding Towards Mngf, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/igg1 hum αd11 binding towards mngf/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    igg1 hum αd11 binding towards mngf - by Bioz Stars, 2023-01
    94/100 stars

    Images

    1) Product Images from "Single Cycle Structure-Based Humanization of an Anti-Nerve Growth Factor Therapeutic Antibody"

    Article Title: Single Cycle Structure-Based Humanization of an Anti-Nerve Growth Factor Therapeutic Antibody

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0032212

    (a) Sequence alignment of mNGF, hNGF and rat NGF (Loop I and Loop II are colored in magenta and green, respectively). (b) Overall structure of NGF and structural comparisons , of Loop I (in magenta) and Loop II (in green) from mNGF (PDB_ID: 1BTG, cyan) and hNGF (PDB_ID: 1WWW, green) . This Figure was prepared with PyMOL . (c) ELISA assay with mNGF and hNGF coating (5 µg/ml) and serial dilutions of parental mAb αD11. The experiments were done in duplicate.
    Figure Legend Snippet: (a) Sequence alignment of mNGF, hNGF and rat NGF (Loop I and Loop II are colored in magenta and green, respectively). (b) Overall structure of NGF and structural comparisons , of Loop I (in magenta) and Loop II (in green) from mNGF (PDB_ID: 1BTG, cyan) and hNGF (PDB_ID: 1WWW, green) . This Figure was prepared with PyMOL . (c) ELISA assay with mNGF and hNGF coating (5 µg/ml) and serial dilutions of parental mAb αD11. The experiments were done in duplicate.

    Techniques Used: Sequencing, Enzyme-linked Immunosorbent Assay

    (a) Analgesic effects of αD11 antibody administration on the late phase (15–40 min) in the course of the formalin test. Treatment consisted in saline (negative control) or antibody injection (single doses: 12.5 µg of mock mouse mAb or two different molecular formats of αD11, i.e. mAb or Fab) performed (in the same paw as for formalin) 45 min before formalin injection and testing. Statistical analysis was performed on each phase (ANOVA and Fisher's Test for comparison of each couple of groups). Each experimental group included at least 8 animals. (b) Analgesic effects of αD11 antibody in the short lasting protocol of neuropathic pain model: mAb αD11 significantly increased the value of ipsilateral/contralateral index (ratio between the threshold forces measured for the two hind paws, the one ipsilateral to surgery and the contralateral one. Mean value ± s.e.), starting from day 4 to day 14, one week after the last antibody injection. Control mice were injected with either mock mouse IgG, (1.4 mg/Kg) or saline solution (sal). ANOVA test for repeated measures resulted in statistical significance for treatment (p<0.0001), time (p<0.0001) and the interaction between the two factors (treatment×time) (p<0.0001). (c) Analgesic efficacy of mAb αD11 (one dose: 2 mg/kg) in the long lasting protocol of neuropathic pain model. MAb αD11 increased the ipsilateral/contralateral index, starting either from day 5. The analgesic effect, which disappeared around days 17–19, increases again to reach a plateau between day 27 and day 31, identifying a late phase in the action of mAb αD11 (long-term effect). ANOVA test for repeated measures resulted in statistical significance for treatment (p<0.005), time (p<0.005) and the interaction between two factors (treatment×time) (p<0.005).
    Figure Legend Snippet: (a) Analgesic effects of αD11 antibody administration on the late phase (15–40 min) in the course of the formalin test. Treatment consisted in saline (negative control) or antibody injection (single doses: 12.5 µg of mock mouse mAb or two different molecular formats of αD11, i.e. mAb or Fab) performed (in the same paw as for formalin) 45 min before formalin injection and testing. Statistical analysis was performed on each phase (ANOVA and Fisher's Test for comparison of each couple of groups). Each experimental group included at least 8 animals. (b) Analgesic effects of αD11 antibody in the short lasting protocol of neuropathic pain model: mAb αD11 significantly increased the value of ipsilateral/contralateral index (ratio between the threshold forces measured for the two hind paws, the one ipsilateral to surgery and the contralateral one. Mean value ± s.e.), starting from day 4 to day 14, one week after the last antibody injection. Control mice were injected with either mock mouse IgG, (1.4 mg/Kg) or saline solution (sal). ANOVA test for repeated measures resulted in statistical significance for treatment (p<0.0001), time (p<0.0001) and the interaction between the two factors (treatment×time) (p<0.0001). (c) Analgesic efficacy of mAb αD11 (one dose: 2 mg/kg) in the long lasting protocol of neuropathic pain model. MAb αD11 increased the ipsilateral/contralateral index, starting either from day 5. The analgesic effect, which disappeared around days 17–19, increases again to reach a plateau between day 27 and day 31, identifying a late phase in the action of mAb αD11 (long-term effect). ANOVA test for repeated measures resulted in statistical significance for treatment (p<0.005), time (p<0.005) and the interaction between two factors (treatment×time) (p<0.005).

    Techniques Used: Negative Control, Injection

    Deviations of parental Fab αD11 from each of the selected human and humanized antibody, taking into account the sequence alignment and structural comparison (calculated considering both the overall sequence homology and identity percentage and the percentage of sequence homology and identity restricted to FWRs).
    Figure Legend Snippet: Deviations of parental Fab αD11 from each of the selected human and humanized antibody, taking into account the sequence alignment and structural comparison (calculated considering both the overall sequence homology and identity percentage and the percentage of sequence homology and identity restricted to FWRs).

    Techniques Used: Sequencing

    Sequence alignment of V κ (A) and V H (B), respectively of parental Fab αD11 (PDB_ID: 1ZAN) , with the selected template for humanization (PDB_ID: 1JPS) and hum-αD11, obtained by CDRs grafting (highlighted in yellow) on PDB_ID: 1JPS FWRs (highlighted in magenta) with the retro-mutations (highlighted in green) and the mutation (highlighted in cyan) . The six CDRs are underlined and the residues belonging to the Vernier zones are colored in red. The residues numbering is according to Kabat .
    Figure Legend Snippet: Sequence alignment of V κ (A) and V H (B), respectively of parental Fab αD11 (PDB_ID: 1ZAN) , with the selected template for humanization (PDB_ID: 1JPS) and hum-αD11, obtained by CDRs grafting (highlighted in yellow) on PDB_ID: 1JPS FWRs (highlighted in magenta) with the retro-mutations (highlighted in green) and the mutation (highlighted in cyan) . The six CDRs are underlined and the residues belonging to the Vernier zones are colored in red. The residues numbering is according to Kabat .

    Techniques Used: Sequencing, Mutagenesis

    Structural alignment between the variable domains of parental Fab αD11 (PDB_ID. 1ZAN) , in magenta (a) with the selected FWRs acceptor 1JPS (in cyan) and (b) with the model of the resulting humanized antibody after CDRs grafting (whose FWRs are depicted in cyan, while its CDRs are in magenta); (c) Model of the hum-αD11 after CDR grafting and mutagenesis in the chosen FWRs. The residues of human and animal origin are highlighted in cyan and violet, respectively.
    Figure Legend Snippet: Structural alignment between the variable domains of parental Fab αD11 (PDB_ID. 1ZAN) , in magenta (a) with the selected FWRs acceptor 1JPS (in cyan) and (b) with the model of the resulting humanized antibody after CDRs grafting (whose FWRs are depicted in cyan, while its CDRs are in magenta); (c) Model of the hum-αD11 after CDR grafting and mutagenesis in the chosen FWRs. The residues of human and animal origin are highlighted in cyan and violet, respectively.

    Techniques Used: Mutagenesis

    (a) ELISA assay with mNGF coating (5 µg/ml); serial dilutions of parental mAb αD11, chimeric IgG1 αD11 and IgG1 hum-αD11, Protein A-Sepharose purified from transiently cotransfected CHO cells supernatants. (b) Binding curves of a range of concentrations (0.29–37.5 nM) of hNGF to immobilized parental mAb αD11 (immobilization level 1280.0 RU). (c) Binding curves of a range of concentrations (0.39–25.0 nM) of Fab hum-αD11 to immobilized hNGF (immobilization level 100.3 RU).
    Figure Legend Snippet: (a) ELISA assay with mNGF coating (5 µg/ml); serial dilutions of parental mAb αD11, chimeric IgG1 αD11 and IgG1 hum-αD11, Protein A-Sepharose purified from transiently cotransfected CHO cells supernatants. (b) Binding curves of a range of concentrations (0.29–37.5 nM) of hNGF to immobilized parental mAb αD11 (immobilization level 1280.0 RU). (c) Binding curves of a range of concentrations (0.39–25.0 nM) of Fab hum-αD11 to immobilized hNGF (immobilization level 100.3 RU).

    Techniques Used: Enzyme-linked Immunosorbent Assay, Purification, Binding Assay

    SPR analysis.
    Figure Legend Snippet: SPR analysis.

    Techniques Used:

    (a–d) mNGF induced differentiation of PC-12 cells. Neurite outgrowth inhibition assay: photomicrographs of PC-12 cells, treated with (a) mNGF 100 ng/ml alone or preincubated (b) with mAb αD11 (5 µg/ml) or (c) with IgG1 hum-αD11 (5 µg/ml), concentrated from stable cotransfected CHO cells supernatants. (d) Negative control: untreated PC-12 cells. (e) TrkA phosphorylation inhibition assay in 3T3 TrkA cells. 3T3 TrkA cells were incubated with the indicated combinations of mNGF, parental mAb αD11, IgG1 hum-αD11 (purified by Protein A-Sepharose) and the irrelevant mAb SV5 as a negative control. Cell lysates were separated on a 10% SDS gel and phosphorylated TrkA detected using anti-phospho-Y490 TrkA Ab. The ubiquitous band of tubulin served as gel loading control.
    Figure Legend Snippet: (a–d) mNGF induced differentiation of PC-12 cells. Neurite outgrowth inhibition assay: photomicrographs of PC-12 cells, treated with (a) mNGF 100 ng/ml alone or preincubated (b) with mAb αD11 (5 µg/ml) or (c) with IgG1 hum-αD11 (5 µg/ml), concentrated from stable cotransfected CHO cells supernatants. (d) Negative control: untreated PC-12 cells. (e) TrkA phosphorylation inhibition assay in 3T3 TrkA cells. 3T3 TrkA cells were incubated with the indicated combinations of mNGF, parental mAb αD11, IgG1 hum-αD11 (purified by Protein A-Sepharose) and the irrelevant mAb SV5 as a negative control. Cell lysates were separated on a 10% SDS gel and phosphorylated TrkA detected using anti-phospho-Y490 TrkA Ab. The ubiquitous band of tubulin served as gel loading control.

    Techniques Used: Inhibition, Negative Control, Incubation, Purification, SDS-Gel

    Dose/Response effect of Fab hum-αD11 on the early (0–15 min) and late phase (15–40 min) in the course of the formalin test in CD1 mice. Treatment consisted in antibody injection (Fab hum-αD11 vs mock IgG) performed (in the same paw as for formalin) 45 min, before formalin injection and testing. Statistical analysis was performed on each phase (ANOVA and Fisher's Test for comparison of each couple of groups).
    Figure Legend Snippet: Dose/Response effect of Fab hum-αD11 on the early (0–15 min) and late phase (15–40 min) in the course of the formalin test in CD1 mice. Treatment consisted in antibody injection (Fab hum-αD11 vs mock IgG) performed (in the same paw as for formalin) 45 min, before formalin injection and testing. Statistical analysis was performed on each phase (ANOVA and Fisher's Test for comparison of each couple of groups).

    Techniques Used: Injection

    mngf  (Alomone Labs)


    Bioz Verified Symbol Alomone Labs is a verified supplier
    Bioz Manufacturer Symbol Alomone Labs manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94

    Structured Review

    Alomone Labs mngf
    Mngf, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mngf/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mngf - by Bioz Stars, 2023-01
    94/100 stars

    Images

    mngf 2 5  (Alomone Labs)


    Bioz Verified Symbol Alomone Labs is a verified supplier
    Bioz Manufacturer Symbol Alomone Labs manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 95

    Structured Review

    Alomone Labs mngf 2 5
    Mngf 2 5, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mngf 2 5/product/Alomone Labs
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mngf 2 5 - by Bioz Stars, 2023-01
    95/100 stars

    Images

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94
    Alomone Labs mngf
    Mngf, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mngf/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mngf - by Bioz Stars, 2023-01
    94/100 stars
      Buy from Supplier

    94
    Alomone Labs igg1 hum αd11 binding towards mngf
    (a) Sequence alignment of <t>mNGF,</t> hNGF and rat NGF (Loop I and Loop II are colored in magenta and green, respectively). (b) Overall structure of NGF and structural comparisons , of Loop I (in magenta) and Loop II (in green) from mNGF (PDB_ID: 1BTG, cyan) and hNGF (PDB_ID: 1WWW, green) . This Figure was prepared with PyMOL . (c) ELISA assay with mNGF and hNGF coating (5 µg/ml) and serial dilutions of parental mAb <t>αD11.</t> The experiments were done in duplicate.
    Igg1 Hum αd11 Binding Towards Mngf, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/igg1 hum αd11 binding towards mngf/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    igg1 hum αd11 binding towards mngf - by Bioz Stars, 2023-01
    94/100 stars
      Buy from Supplier

    95
    Alomone Labs mngf 2 5
    (a) Sequence alignment of <t>mNGF,</t> hNGF and rat NGF (Loop I and Loop II are colored in magenta and green, respectively). (b) Overall structure of NGF and structural comparisons , of Loop I (in magenta) and Loop II (in green) from mNGF (PDB_ID: 1BTG, cyan) and hNGF (PDB_ID: 1WWW, green) . This Figure was prepared with PyMOL . (c) ELISA assay with mNGF and hNGF coating (5 µg/ml) and serial dilutions of parental mAb <t>αD11.</t> The experiments were done in duplicate.
    Mngf 2 5, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mngf 2 5/product/Alomone Labs
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mngf 2 5 - by Bioz Stars, 2023-01
    95/100 stars
      Buy from Supplier

    Image Search Results


    (a) Sequence alignment of mNGF, hNGF and rat NGF (Loop I and Loop II are colored in magenta and green, respectively). (b) Overall structure of NGF and structural comparisons , of Loop I (in magenta) and Loop II (in green) from mNGF (PDB_ID: 1BTG, cyan) and hNGF (PDB_ID: 1WWW, green) . This Figure was prepared with PyMOL . (c) ELISA assay with mNGF and hNGF coating (5 µg/ml) and serial dilutions of parental mAb αD11. The experiments were done in duplicate.

    Journal: PLoS ONE

    Article Title: Single Cycle Structure-Based Humanization of an Anti-Nerve Growth Factor Therapeutic Antibody

    doi: 10.1371/journal.pone.0032212

    Figure Lengend Snippet: (a) Sequence alignment of mNGF, hNGF and rat NGF (Loop I and Loop II are colored in magenta and green, respectively). (b) Overall structure of NGF and structural comparisons , of Loop I (in magenta) and Loop II (in green) from mNGF (PDB_ID: 1BTG, cyan) and hNGF (PDB_ID: 1WWW, green) . This Figure was prepared with PyMOL . (c) ELISA assay with mNGF and hNGF coating (5 µg/ml) and serial dilutions of parental mAb αD11. The experiments were done in duplicate.

    Article Snippet: ELISA assay to compare chimeric and IgG1 hum-αD11 binding towards mNGF (Alomone) was performed with the following modifications: serial dilutions of chimeric and IgG1 hum-αD11 were incubated and the anti-human polyclonal antibody (Pierce) was used as primary antibody and the anti-goat antibody peroxidase conjugated as secondary antibody (Dako).

    Techniques: Sequencing, Enzyme-linked Immunosorbent Assay

    (a) Analgesic effects of αD11 antibody administration on the late phase (15–40 min) in the course of the formalin test. Treatment consisted in saline (negative control) or antibody injection (single doses: 12.5 µg of mock mouse mAb or two different molecular formats of αD11, i.e. mAb or Fab) performed (in the same paw as for formalin) 45 min before formalin injection and testing. Statistical analysis was performed on each phase (ANOVA and Fisher's Test for comparison of each couple of groups). Each experimental group included at least 8 animals. (b) Analgesic effects of αD11 antibody in the short lasting protocol of neuropathic pain model: mAb αD11 significantly increased the value of ipsilateral/contralateral index (ratio between the threshold forces measured for the two hind paws, the one ipsilateral to surgery and the contralateral one. Mean value ± s.e.), starting from day 4 to day 14, one week after the last antibody injection. Control mice were injected with either mock mouse IgG, (1.4 mg/Kg) or saline solution (sal). ANOVA test for repeated measures resulted in statistical significance for treatment (p<0.0001), time (p<0.0001) and the interaction between the two factors (treatment×time) (p<0.0001). (c) Analgesic efficacy of mAb αD11 (one dose: 2 mg/kg) in the long lasting protocol of neuropathic pain model. MAb αD11 increased the ipsilateral/contralateral index, starting either from day 5. The analgesic effect, which disappeared around days 17–19, increases again to reach a plateau between day 27 and day 31, identifying a late phase in the action of mAb αD11 (long-term effect). ANOVA test for repeated measures resulted in statistical significance for treatment (p<0.005), time (p<0.005) and the interaction between two factors (treatment×time) (p<0.005).

    Journal: PLoS ONE

    Article Title: Single Cycle Structure-Based Humanization of an Anti-Nerve Growth Factor Therapeutic Antibody

    doi: 10.1371/journal.pone.0032212

    Figure Lengend Snippet: (a) Analgesic effects of αD11 antibody administration on the late phase (15–40 min) in the course of the formalin test. Treatment consisted in saline (negative control) or antibody injection (single doses: 12.5 µg of mock mouse mAb or two different molecular formats of αD11, i.e. mAb or Fab) performed (in the same paw as for formalin) 45 min before formalin injection and testing. Statistical analysis was performed on each phase (ANOVA and Fisher's Test for comparison of each couple of groups). Each experimental group included at least 8 animals. (b) Analgesic effects of αD11 antibody in the short lasting protocol of neuropathic pain model: mAb αD11 significantly increased the value of ipsilateral/contralateral index (ratio between the threshold forces measured for the two hind paws, the one ipsilateral to surgery and the contralateral one. Mean value ± s.e.), starting from day 4 to day 14, one week after the last antibody injection. Control mice were injected with either mock mouse IgG, (1.4 mg/Kg) or saline solution (sal). ANOVA test for repeated measures resulted in statistical significance for treatment (p<0.0001), time (p<0.0001) and the interaction between the two factors (treatment×time) (p<0.0001). (c) Analgesic efficacy of mAb αD11 (one dose: 2 mg/kg) in the long lasting protocol of neuropathic pain model. MAb αD11 increased the ipsilateral/contralateral index, starting either from day 5. The analgesic effect, which disappeared around days 17–19, increases again to reach a plateau between day 27 and day 31, identifying a late phase in the action of mAb αD11 (long-term effect). ANOVA test for repeated measures resulted in statistical significance for treatment (p<0.005), time (p<0.005) and the interaction between two factors (treatment×time) (p<0.005).

    Article Snippet: ELISA assay to compare chimeric and IgG1 hum-αD11 binding towards mNGF (Alomone) was performed with the following modifications: serial dilutions of chimeric and IgG1 hum-αD11 were incubated and the anti-human polyclonal antibody (Pierce) was used as primary antibody and the anti-goat antibody peroxidase conjugated as secondary antibody (Dako).

    Techniques: Negative Control, Injection

    Deviations of parental Fab αD11 from each of the selected human and humanized antibody, taking into account the sequence alignment and structural comparison (calculated considering both the overall sequence homology and identity percentage and the percentage of sequence homology and identity restricted to FWRs).

    Journal: PLoS ONE

    Article Title: Single Cycle Structure-Based Humanization of an Anti-Nerve Growth Factor Therapeutic Antibody

    doi: 10.1371/journal.pone.0032212

    Figure Lengend Snippet: Deviations of parental Fab αD11 from each of the selected human and humanized antibody, taking into account the sequence alignment and structural comparison (calculated considering both the overall sequence homology and identity percentage and the percentage of sequence homology and identity restricted to FWRs).

    Article Snippet: ELISA assay to compare chimeric and IgG1 hum-αD11 binding towards mNGF (Alomone) was performed with the following modifications: serial dilutions of chimeric and IgG1 hum-αD11 were incubated and the anti-human polyclonal antibody (Pierce) was used as primary antibody and the anti-goat antibody peroxidase conjugated as secondary antibody (Dako).

    Techniques: Sequencing

    Sequence alignment of V κ (A) and V H (B), respectively of parental Fab αD11 (PDB_ID: 1ZAN) , with the selected template for humanization (PDB_ID: 1JPS) and hum-αD11, obtained by CDRs grafting (highlighted in yellow) on PDB_ID: 1JPS FWRs (highlighted in magenta) with the retro-mutations (highlighted in green) and the mutation (highlighted in cyan) . The six CDRs are underlined and the residues belonging to the Vernier zones are colored in red. The residues numbering is according to Kabat .

    Journal: PLoS ONE

    Article Title: Single Cycle Structure-Based Humanization of an Anti-Nerve Growth Factor Therapeutic Antibody

    doi: 10.1371/journal.pone.0032212

    Figure Lengend Snippet: Sequence alignment of V κ (A) and V H (B), respectively of parental Fab αD11 (PDB_ID: 1ZAN) , with the selected template for humanization (PDB_ID: 1JPS) and hum-αD11, obtained by CDRs grafting (highlighted in yellow) on PDB_ID: 1JPS FWRs (highlighted in magenta) with the retro-mutations (highlighted in green) and the mutation (highlighted in cyan) . The six CDRs are underlined and the residues belonging to the Vernier zones are colored in red. The residues numbering is according to Kabat .

    Article Snippet: ELISA assay to compare chimeric and IgG1 hum-αD11 binding towards mNGF (Alomone) was performed with the following modifications: serial dilutions of chimeric and IgG1 hum-αD11 were incubated and the anti-human polyclonal antibody (Pierce) was used as primary antibody and the anti-goat antibody peroxidase conjugated as secondary antibody (Dako).

    Techniques: Sequencing, Mutagenesis

    Structural alignment between the variable domains of parental Fab αD11 (PDB_ID. 1ZAN) , in magenta (a) with the selected FWRs acceptor 1JPS (in cyan) and (b) with the model of the resulting humanized antibody after CDRs grafting (whose FWRs are depicted in cyan, while its CDRs are in magenta); (c) Model of the hum-αD11 after CDR grafting and mutagenesis in the chosen FWRs. The residues of human and animal origin are highlighted in cyan and violet, respectively.

    Journal: PLoS ONE

    Article Title: Single Cycle Structure-Based Humanization of an Anti-Nerve Growth Factor Therapeutic Antibody

    doi: 10.1371/journal.pone.0032212

    Figure Lengend Snippet: Structural alignment between the variable domains of parental Fab αD11 (PDB_ID. 1ZAN) , in magenta (a) with the selected FWRs acceptor 1JPS (in cyan) and (b) with the model of the resulting humanized antibody after CDRs grafting (whose FWRs are depicted in cyan, while its CDRs are in magenta); (c) Model of the hum-αD11 after CDR grafting and mutagenesis in the chosen FWRs. The residues of human and animal origin are highlighted in cyan and violet, respectively.

    Article Snippet: ELISA assay to compare chimeric and IgG1 hum-αD11 binding towards mNGF (Alomone) was performed with the following modifications: serial dilutions of chimeric and IgG1 hum-αD11 were incubated and the anti-human polyclonal antibody (Pierce) was used as primary antibody and the anti-goat antibody peroxidase conjugated as secondary antibody (Dako).

    Techniques: Mutagenesis

    (a) ELISA assay with mNGF coating (5 µg/ml); serial dilutions of parental mAb αD11, chimeric IgG1 αD11 and IgG1 hum-αD11, Protein A-Sepharose purified from transiently cotransfected CHO cells supernatants. (b) Binding curves of a range of concentrations (0.29–37.5 nM) of hNGF to immobilized parental mAb αD11 (immobilization level 1280.0 RU). (c) Binding curves of a range of concentrations (0.39–25.0 nM) of Fab hum-αD11 to immobilized hNGF (immobilization level 100.3 RU).

    Journal: PLoS ONE

    Article Title: Single Cycle Structure-Based Humanization of an Anti-Nerve Growth Factor Therapeutic Antibody

    doi: 10.1371/journal.pone.0032212

    Figure Lengend Snippet: (a) ELISA assay with mNGF coating (5 µg/ml); serial dilutions of parental mAb αD11, chimeric IgG1 αD11 and IgG1 hum-αD11, Protein A-Sepharose purified from transiently cotransfected CHO cells supernatants. (b) Binding curves of a range of concentrations (0.29–37.5 nM) of hNGF to immobilized parental mAb αD11 (immobilization level 1280.0 RU). (c) Binding curves of a range of concentrations (0.39–25.0 nM) of Fab hum-αD11 to immobilized hNGF (immobilization level 100.3 RU).

    Article Snippet: ELISA assay to compare chimeric and IgG1 hum-αD11 binding towards mNGF (Alomone) was performed with the following modifications: serial dilutions of chimeric and IgG1 hum-αD11 were incubated and the anti-human polyclonal antibody (Pierce) was used as primary antibody and the anti-goat antibody peroxidase conjugated as secondary antibody (Dako).

    Techniques: Enzyme-linked Immunosorbent Assay, Purification, Binding Assay

    SPR analysis.

    Journal: PLoS ONE

    Article Title: Single Cycle Structure-Based Humanization of an Anti-Nerve Growth Factor Therapeutic Antibody

    doi: 10.1371/journal.pone.0032212

    Figure Lengend Snippet: SPR analysis.

    Article Snippet: ELISA assay to compare chimeric and IgG1 hum-αD11 binding towards mNGF (Alomone) was performed with the following modifications: serial dilutions of chimeric and IgG1 hum-αD11 were incubated and the anti-human polyclonal antibody (Pierce) was used as primary antibody and the anti-goat antibody peroxidase conjugated as secondary antibody (Dako).

    Techniques:

    (a–d) mNGF induced differentiation of PC-12 cells. Neurite outgrowth inhibition assay: photomicrographs of PC-12 cells, treated with (a) mNGF 100 ng/ml alone or preincubated (b) with mAb αD11 (5 µg/ml) or (c) with IgG1 hum-αD11 (5 µg/ml), concentrated from stable cotransfected CHO cells supernatants. (d) Negative control: untreated PC-12 cells. (e) TrkA phosphorylation inhibition assay in 3T3 TrkA cells. 3T3 TrkA cells were incubated with the indicated combinations of mNGF, parental mAb αD11, IgG1 hum-αD11 (purified by Protein A-Sepharose) and the irrelevant mAb SV5 as a negative control. Cell lysates were separated on a 10% SDS gel and phosphorylated TrkA detected using anti-phospho-Y490 TrkA Ab. The ubiquitous band of tubulin served as gel loading control.

    Journal: PLoS ONE

    Article Title: Single Cycle Structure-Based Humanization of an Anti-Nerve Growth Factor Therapeutic Antibody

    doi: 10.1371/journal.pone.0032212

    Figure Lengend Snippet: (a–d) mNGF induced differentiation of PC-12 cells. Neurite outgrowth inhibition assay: photomicrographs of PC-12 cells, treated with (a) mNGF 100 ng/ml alone or preincubated (b) with mAb αD11 (5 µg/ml) or (c) with IgG1 hum-αD11 (5 µg/ml), concentrated from stable cotransfected CHO cells supernatants. (d) Negative control: untreated PC-12 cells. (e) TrkA phosphorylation inhibition assay in 3T3 TrkA cells. 3T3 TrkA cells were incubated with the indicated combinations of mNGF, parental mAb αD11, IgG1 hum-αD11 (purified by Protein A-Sepharose) and the irrelevant mAb SV5 as a negative control. Cell lysates were separated on a 10% SDS gel and phosphorylated TrkA detected using anti-phospho-Y490 TrkA Ab. The ubiquitous band of tubulin served as gel loading control.

    Article Snippet: ELISA assay to compare chimeric and IgG1 hum-αD11 binding towards mNGF (Alomone) was performed with the following modifications: serial dilutions of chimeric and IgG1 hum-αD11 were incubated and the anti-human polyclonal antibody (Pierce) was used as primary antibody and the anti-goat antibody peroxidase conjugated as secondary antibody (Dako).

    Techniques: Inhibition, Negative Control, Incubation, Purification, SDS-Gel

    Dose/Response effect of Fab hum-αD11 on the early (0–15 min) and late phase (15–40 min) in the course of the formalin test in CD1 mice. Treatment consisted in antibody injection (Fab hum-αD11 vs mock IgG) performed (in the same paw as for formalin) 45 min, before formalin injection and testing. Statistical analysis was performed on each phase (ANOVA and Fisher's Test for comparison of each couple of groups).

    Journal: PLoS ONE

    Article Title: Single Cycle Structure-Based Humanization of an Anti-Nerve Growth Factor Therapeutic Antibody

    doi: 10.1371/journal.pone.0032212

    Figure Lengend Snippet: Dose/Response effect of Fab hum-αD11 on the early (0–15 min) and late phase (15–40 min) in the course of the formalin test in CD1 mice. Treatment consisted in antibody injection (Fab hum-αD11 vs mock IgG) performed (in the same paw as for formalin) 45 min, before formalin injection and testing. Statistical analysis was performed on each phase (ANOVA and Fisher's Test for comparison of each couple of groups).

    Article Snippet: ELISA assay to compare chimeric and IgG1 hum-αD11 binding towards mNGF (Alomone) was performed with the following modifications: serial dilutions of chimeric and IgG1 hum-αD11 were incubated and the anti-human polyclonal antibody (Pierce) was used as primary antibody and the anti-goat antibody peroxidase conjugated as secondary antibody (Dako).

    Techniques: Injection