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Sino Biological recombinant murine
Binding of selected <t>recombinant</t> anti-CAIX antibodies by Surface Plasmon Resonance. Six (6) hybridoma derived mAbs were selected through screening (see , Suppl. Fig. S2A-C), sequenced, recloned in the appropriate IgG framework and recombinantly expressed in CHO cells (for details see text). (a) Purified recombinant antibodies (c11H9, c12H8, c2C7, m4A2, m9B6, c2D7) were captured with the appropriate anti-Fc surface (anti-human Fc: c11H9, c12H8, c2C7, c2D7; anti-mouse Fc: m4A2, m9B6). cG250 was used as a benchmark. Serial dilutions (0.74–60 nM) of rhCAIX monomer (CAIX-M) and dimer (CAIX-D) were then injected, followed by a buffer injection. Sensorgrams were aligned, double-referenced, and fitted to the 1:1 binding model to calculate ka, kd, KD and RUmax when flowing CAIX-M and apparent ka, kd, KD and RUmax when flowing CAIX-D (see ). (b) Graphs depicting changes in the calculated ka, kd, KD and RUmax when flowing rhCAIX-M versus rhCAIX-D over the immobilized antibodies
Recombinant Murine, supplied by Sino Biological, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Binding of selected recombinant anti-CAIX antibodies by Surface Plasmon Resonance. Six (6) hybridoma derived mAbs were selected through screening (see , Suppl. Fig. S2A-C), sequenced, recloned in the appropriate IgG framework and recombinantly expressed in CHO cells (for details see text). (a) Purified recombinant antibodies (c11H9, c12H8, c2C7, m4A2, m9B6, c2D7) were captured with the appropriate anti-Fc surface (anti-human Fc: c11H9, c12H8, c2C7, c2D7; anti-mouse Fc: m4A2, m9B6). cG250 was used as a benchmark. Serial dilutions (0.74–60 nM) of rhCAIX monomer (CAIX-M) and dimer (CAIX-D) were then injected, followed by a buffer injection. Sensorgrams were aligned, double-referenced, and fitted to the 1:1 binding model to calculate ka, kd, KD and RUmax when flowing CAIX-M and apparent ka, kd, KD and RUmax when flowing CAIX-D (see ). (b) Graphs depicting changes in the calculated ka, kd, KD and RUmax when flowing rhCAIX-M versus rhCAIX-D over the immobilized antibodies

Journal: mAbs

Article Title: Isolation and characterization of monoclonal antibodies against human carbonic anhydrase-IX

doi: 10.1080/19420862.2021.1999194

Figure Lengend Snippet: Binding of selected recombinant anti-CAIX antibodies by Surface Plasmon Resonance. Six (6) hybridoma derived mAbs were selected through screening (see , Suppl. Fig. S2A-C), sequenced, recloned in the appropriate IgG framework and recombinantly expressed in CHO cells (for details see text). (a) Purified recombinant antibodies (c11H9, c12H8, c2C7, m4A2, m9B6, c2D7) were captured with the appropriate anti-Fc surface (anti-human Fc: c11H9, c12H8, c2C7, c2D7; anti-mouse Fc: m4A2, m9B6). cG250 was used as a benchmark. Serial dilutions (0.74–60 nM) of rhCAIX monomer (CAIX-M) and dimer (CAIX-D) were then injected, followed by a buffer injection. Sensorgrams were aligned, double-referenced, and fitted to the 1:1 binding model to calculate ka, kd, KD and RUmax when flowing CAIX-M and apparent ka, kd, KD and RUmax when flowing CAIX-D (see ). (b) Graphs depicting changes in the calculated ka, kd, KD and RUmax when flowing rhCAIX-M versus rhCAIX-D over the immobilized antibodies

Article Snippet: Mouse monoclonal anti-human CAIX MAB2188 was obtained from R&D Systems (Cat# MAB2188), goat anti-human and goat anti-mouse AlexaFluor(AF)488-labeleled IgG-Fab2 were purchased from Jackson ImmunoResearch (Cat# 109–547-003 and 115–547-003, respectively), recombinant human (rh)CAIV (Cat# 510472-H08S), CAXII (Cat# 10617-H08H) and CAXIV (Cat# 10458H08H) and recombinant murine (rm; Cat# 50660-M08H) and dog (rd; Cat# 70028D08H) CAIX were purchased from Sino Biologics Inc., and cynomolgus CAIX was purchased from ACRO Biosystems (Cat# CA9-C83E6).

Techniques: Binding Assay, Recombinant, SPR Assay, Derivative Assay, Purification, Injection

Representative kinetic and apparent kinetic values for binding of recombinant anti-CAIX antibodies obtained by SPR analysis and by flowing rhCAIX monomer or dimer, respectively. Antibodies suitable for ADC, CAIX enzyme inhibitor and imaging/detection are listed in green, blue and red, respectively. The cG250 mAb was used as control (black)

Journal: mAbs

Article Title: Isolation and characterization of monoclonal antibodies against human carbonic anhydrase-IX

doi: 10.1080/19420862.2021.1999194

Figure Lengend Snippet: Representative kinetic and apparent kinetic values for binding of recombinant anti-CAIX antibodies obtained by SPR analysis and by flowing rhCAIX monomer or dimer, respectively. Antibodies suitable for ADC, CAIX enzyme inhibitor and imaging/detection are listed in green, blue and red, respectively. The cG250 mAb was used as control (black)

Article Snippet: Mouse monoclonal anti-human CAIX MAB2188 was obtained from R&D Systems (Cat# MAB2188), goat anti-human and goat anti-mouse AlexaFluor(AF)488-labeleled IgG-Fab2 were purchased from Jackson ImmunoResearch (Cat# 109–547-003 and 115–547-003, respectively), recombinant human (rh)CAIV (Cat# 510472-H08S), CAXII (Cat# 10617-H08H) and CAXIV (Cat# 10458H08H) and recombinant murine (rm; Cat# 50660-M08H) and dog (rd; Cat# 70028D08H) CAIX were purchased from Sino Biologics Inc., and cynomolgus CAIX was purchased from ACRO Biosystems (Cat# CA9-C83E6).

Techniques: Binding Assay, Recombinant, Imaging, Control

Binding of recombinant antibodies to hCAIX-expressing SK-RC-52 cells. Dose-dependent binding (0–100 nM) to SK-RC-52 cells of recombinantly expressed (a) chimeric antibodies (c11H9, c12H8, c2C7, c2D7) and (b) murine antibodies (m4A2, m9B6), together with the appropriate negative (hIgG1, mIgG2) and positive (cG250, M75) controls. Experiments were carried out in duplicate and repeated twice. Representative KD, Hill slope and Bmax (see ) values were calculated using Graphpad Prism v8

Journal: mAbs

Article Title: Isolation and characterization of monoclonal antibodies against human carbonic anhydrase-IX

doi: 10.1080/19420862.2021.1999194

Figure Lengend Snippet: Binding of recombinant antibodies to hCAIX-expressing SK-RC-52 cells. Dose-dependent binding (0–100 nM) to SK-RC-52 cells of recombinantly expressed (a) chimeric antibodies (c11H9, c12H8, c2C7, c2D7) and (b) murine antibodies (m4A2, m9B6), together with the appropriate negative (hIgG1, mIgG2) and positive (cG250, M75) controls. Experiments were carried out in duplicate and repeated twice. Representative KD, Hill slope and Bmax (see ) values were calculated using Graphpad Prism v8

Article Snippet: Mouse monoclonal anti-human CAIX MAB2188 was obtained from R&D Systems (Cat# MAB2188), goat anti-human and goat anti-mouse AlexaFluor(AF)488-labeleled IgG-Fab2 were purchased from Jackson ImmunoResearch (Cat# 109–547-003 and 115–547-003, respectively), recombinant human (rh)CAIV (Cat# 510472-H08S), CAXII (Cat# 10617-H08H) and CAXIV (Cat# 10458H08H) and recombinant murine (rm; Cat# 50660-M08H) and dog (rd; Cat# 70028D08H) CAIX were purchased from Sino Biologics Inc., and cynomolgus CAIX was purchased from ACRO Biosystems (Cat# CA9-C83E6).

Techniques: Binding Assay, Recombinant, Expressing

Apparent equilibrium binding constants (EC50) and maximum specific binding (Bmax) values for the recombinant antibodies using SK-RC-52 cells expressing hCA-IX. cG250 and M75(both in black) were used as positive controls; negative controls (not listed) showed no binding (ADC candidates, green; enzyme inhibitors, blue and imaging/detection antibody, red)

Journal: mAbs

Article Title: Isolation and characterization of monoclonal antibodies against human carbonic anhydrase-IX

doi: 10.1080/19420862.2021.1999194

Figure Lengend Snippet: Apparent equilibrium binding constants (EC50) and maximum specific binding (Bmax) values for the recombinant antibodies using SK-RC-52 cells expressing hCA-IX. cG250 and M75(both in black) were used as positive controls; negative controls (not listed) showed no binding (ADC candidates, green; enzyme inhibitors, blue and imaging/detection antibody, red)

Article Snippet: Mouse monoclonal anti-human CAIX MAB2188 was obtained from R&D Systems (Cat# MAB2188), goat anti-human and goat anti-mouse AlexaFluor(AF)488-labeleled IgG-Fab2 were purchased from Jackson ImmunoResearch (Cat# 109–547-003 and 115–547-003, respectively), recombinant human (rh)CAIV (Cat# 510472-H08S), CAXII (Cat# 10617-H08H) and CAXIV (Cat# 10458H08H) and recombinant murine (rm; Cat# 50660-M08H) and dog (rd; Cat# 70028D08H) CAIX were purchased from Sino Biologics Inc., and cynomolgus CAIX was purchased from ACRO Biosystems (Cat# CA9-C83E6).

Techniques: Binding Assay, Recombinant, Expressing, Imaging

Assessment of recombinant anti-hCAIX antibodies as tools for immunohistochemistry and in vivo imaging applications. (a) Immunohistochemical staining for expression of CAIX in FFPE tissue sections from PK-8 human PDAC xenografts using recombinant antibodies c11H9, m9B6 and c2D7. Commercial anti-CAIX mAb, M75, was used as benchmark-based positive control. Scale bar, 100 μm; inset, 20 μm. Anti-CAIX antibody 11H9 (b) and a (c) control antibody (IgG) were conjugated to the chelator pSCN-Bn-DTPA and radiolabelled with 111 In. Conjugates were administered to NODSCID IL2RKO mice bearing subcutaneous hCAIX-positive HT-29 colorectal cancer xenografts (100 mm 3 ). Uptake, accumulation and retention of the radiolabelled 11H9 and IgG control were monitored 24–168 h post-injection by SPECT/CT imaging (B, C; see also ) (t, tumor; l, lung; s, stomach; h, heart)

Journal: mAbs

Article Title: Isolation and characterization of monoclonal antibodies against human carbonic anhydrase-IX

doi: 10.1080/19420862.2021.1999194

Figure Lengend Snippet: Assessment of recombinant anti-hCAIX antibodies as tools for immunohistochemistry and in vivo imaging applications. (a) Immunohistochemical staining for expression of CAIX in FFPE tissue sections from PK-8 human PDAC xenografts using recombinant antibodies c11H9, m9B6 and c2D7. Commercial anti-CAIX mAb, M75, was used as benchmark-based positive control. Scale bar, 100 μm; inset, 20 μm. Anti-CAIX antibody 11H9 (b) and a (c) control antibody (IgG) were conjugated to the chelator pSCN-Bn-DTPA and radiolabelled with 111 In. Conjugates were administered to NODSCID IL2RKO mice bearing subcutaneous hCAIX-positive HT-29 colorectal cancer xenografts (100 mm 3 ). Uptake, accumulation and retention of the radiolabelled 11H9 and IgG control were monitored 24–168 h post-injection by SPECT/CT imaging (B, C; see also ) (t, tumor; l, lung; s, stomach; h, heart)

Article Snippet: Mouse monoclonal anti-human CAIX MAB2188 was obtained from R&D Systems (Cat# MAB2188), goat anti-human and goat anti-mouse AlexaFluor(AF)488-labeleled IgG-Fab2 were purchased from Jackson ImmunoResearch (Cat# 109–547-003 and 115–547-003, respectively), recombinant human (rh)CAIV (Cat# 510472-H08S), CAXII (Cat# 10617-H08H) and CAXIV (Cat# 10458H08H) and recombinant murine (rm; Cat# 50660-M08H) and dog (rd; Cat# 70028D08H) CAIX were purchased from Sino Biologics Inc., and cynomolgus CAIX was purchased from ACRO Biosystems (Cat# CA9-C83E6).

Techniques: Recombinant, Immunohistochemistry, In Vivo Imaging, Immunohistochemical staining, Staining, Expressing, Positive Control, Control, Injection, Single Photon Emission Computed Tomography, Imaging