Human Anti-IL4 Recombinant Antibody (clone hu19C11) (CAT#: HPAB-0082-YC)

Figure 1 Mutagenesis mapping of hu19C11 CDRs.

To measure the relative antigen binding affinity of hu19C11 Fab variants, binding of serially diluted phage displaying anti-IL4 hu19C11 wild type (wt) or LC CDR alanine mutants to IL4 (A) or to anti-gD antibody (B) coated on ELISA wells was detected by an anti-M13 phage horseradish peroxidase (HRP) conjugate to measure antigen binding and Fab expression, respectively. Single letter codes of amino acids are used. gD is an expression peptide tag fused to the C terminus of the light chain. Anti-gD antibody was directly coated on ELISA wells, whereas IL4 was captured with a non-blocking anti-IL4 antibody coated on ELISA wells. (C) Effects of alanine mutations at individual sites of CDRs were examined by comparing the relative IL4 binding affinity of phage displaying Fab variants by performing assays as (A) and (B). Relative IL4 binding affinity was determined by first fitting the data with a linear regression model then dividing the slope of IL4 binding (vs. phage concentration) with the slope of Fab expression. Low values (below the dotted line) were deemed low IL4 binding affinity relative to hu1911wt indicating disruptive mutations.

Lee, C. V., Koenig, P., & Fuh, G. (2014, May). A two-in-one antibody engineered from a humanized interleukin 4 antibody through mutation in heavy chain complementarity-determining regions. In MAbs (Vol. 6, No. 3, pp. 622-627). Taylor & Francis.

Figure 2 Dual specificity of the selected variants of hu19C11 as IgG.

Antigen binding specificity of selected variants of hu19C11 was assessed as binding of these variants in human IgG1 format at 250nM to target antigen(s) or several irrelevant proteins coated on ELISA wells with detection by anti-Fc antibody HRP conjugates.

Lee, C. V., Koenig, P., & Fuh, G. (2014, May). A two-in-one antibody engineered from a humanized interleukin 4 antibody through mutation in heavy chain complementarity-determining regions. In MAbs (Vol. 6, No. 3, pp. 622-627). Taylor & Francis.