Mouse Anti-CD20 Recombinant Antibody (clone scFv-7) (CAT#: FAMAB-0462WJ)

Figure 1 The indicated scFv (3 or 7) was cloned into pCDNA3.1 containing a leader sequence (amino acids MGGS) for targeted secretion into the media of tissue cultured CHO cells.

(i) Binding was measured on left panel after dilution of the supernatants (1:40 or 1:80) to optimize binding against His-tagged CD20 protein; (ii) right panel the optimized supernatants of scFv-3 and scFv-7 were assayed against biotinylated CD20 peptide as indicated in the materials and methods. Antibody scFv binding to antigen was detected using peroxidase conjugated protein A and resolved with TMB-based ELISA at an OD of 450nm. Controls included media only or PBS.

Jain, S., Aresu, L., Comazzi, S., Shi, J., Worrall, E., Clayton, J., ... & Hupp, T. R. (2016). The development of a recombinant scFv monoclonal antibody targeting canine CD20 for use in comparative medicine. PloS one, 11(2), e0148366.

Figure 2 Bioactivity of affinity purified scFv-3 and scFv-7 from bacteria.

The normalized affinity-purified scFv fractions were assayed for binding to the indicated canine CD20 peptides (peptide 32 and peptide 38), where the core epitope resides (in gold shade). scFv-3:CPG2 was more active than scFv-7:CPG2 on both peptide 32 and peptide 38. The enhanced binding of scFv-3:CPG2 as a bioconjugate is consistent with its enhanced binding when secreted as a single chain from CHO cells and when fused to M13 gIII protein.

Jain, S., Aresu, L., Comazzi, S., Shi, J., Worrall, E., Clayton, J., ... & Hupp, T. R. (2016). The development of a recombinant scFv monoclonal antibody targeting canine CD20 for use in comparative medicine. PloS one, 11(2), e0148366.