Mouse Anti-PA Recombinant Antibody (clone M18) (CAT#: PABW-099)

Figure 1 Effect of panning or FACS sorting on enrichment efficiency at low and high initial M18 purity.

Mixtures of spheroplasts at the indicated M18 purity were panned or sorted against H1299-tet onPAD4 cells. M18 percentage was determined by colony PCR after each round. Mean±SD of three independent experiments is shown; P<0.05; Student's t test, comparison of panning, and FACS.

Qiu, J. K., Jung, S. T., Georgiou, G., & Hang, H. Y. (2010). Enrichment of Escherichia coli spheroplasts displaying scFv antibodies specific for antigens expressed on the human cell surface. Applied microbiology and biotechnology, 88(6), 1385-1391.

Figure 2 A western blot of M18 and variants treated with periodate in the absence and presence of antigen PAD4. a-Flag-HRP was used to detect a Flag-tag on the M18 variants.

The shift in mass in the presence of PAD4 corresponds to the mass of PAD4 antigen (14 kDa), signaling covalent crosslinking between antibody and antigen. Bands were quantified using ImageJ to determine crosslinking efficiencies, which were listed in Table 1. D17L is the only tested position on M18 light chain. The rest of variants were all on heavy chain.

Xu, J., Tack, D., Hughes, R. A., Ellington, A. D., & Gray, J. J. (2014). Structure-based non-canonical amino acid design to covalently crosslink an antibody–antigen complex. Journal of structural biology, 185(2), 215-222.