Journal: mAbs

Article Title: Hydrogen-deuterium exchange mass spectrometry reveals three unique binding responses of mAbs directed to the catalytic domain of hCAIX

doi: 10.1080/19420862.2021.1997072

Figure Lengend Snippet: (a-b) Competitive SPR binding demonstrating that mAbs m4A2 and m9B6 bind to independent epitopes. (c) Analysis of CAIX catalytic activity by 67NR murine breast cancer cells expressing hCAIX incubated with anti-CAIX mAb m4A2 and/or m9B6, followed by addition of buffer and CO 2 -saturated water and monitoring the decrease in pH using the “in-cell” carbonic anhydrase activity assay. Cells incubated in the presence of IgG2b were used as a control. Data show the mean ± SEM of technical replicates (n = 3/group) and are representative of 2 independent experiments

Article Snippet: Anti-human CAIX mouse mAbs were sequenced and full-size heavy and light human IgG chains were ordered from a commercial vendor (Genscript); V H and V L CDR1-3 regions of 2C7 was grafted on a human IgG1 framework (designated chimeric, ‘c’) whereas those of 4A2 and 9B6 were grafted on a murine IgG2a framework (designated as mouse, ‘m’).

Techniques: Binding Assay, Activity Assay, Expressing, Incubation, Control