Recombinant Human Anti-Hepatitis B virus Antibody scFv Fragment (KR127)

Figure 1 Validation of epitope specificity with HzKR127-3.2.

GST-preS1 (amino acids 1–56) protein (WT) and a series of alanine replacement mutants were expressed in E. coli and subjected to 12% SDS-PAGE (bottom) followed by western blot analysis with KR127, HzKR127-3.1, or HzKR127-3.2 (Top). The protein band corresponding to the GST-preS1 is marked.

Kim, J. H., Gripon, P., Bouezzedine, F., Jeong, M. S., Chi, S. W., Ryu, S. E., & Hong, H. J. (2015). Enhanced humanization and affinity maturation of neutralizing anti-hepatitis B virus preS1 antibody based on antigen–antibody complex structure. FEBS letters, 589(2), 193-200.

Figure 2 Validation of off-target activity with HzKR127-3.2.

HzKR127-3.2 was subjected to flow cytometric analysis using preS1-expressing HEK293T cells (293T-S1-L1) and preS1-negative cells (HEK293T, SCK-L1, ACHN, B16F1, and CHO).

Kim, J. H., Gripon, P., Bouezzedine, F., Jeong, M. S., Chi, S. W., Ryu, S. E., & Hong, H. J. (2015). Enhanced humanization and affinity maturation of neutralizing anti-hepatitis B virus preS1 antibody based on antigen–antibody complex structure. FEBS letters, 589(2), 193-200.

Figure 3 Affinity determination of murine and humanized KR127 antibody by a competitive ELISA.

A solution containing 3 ng of the purified antibody and various concentrations (1011–106 M) of the GST-preS1 protein as a competing antigen was preincubated at 37 jC for 3 h and added to each well that was coated with 0.1 Ag of the GST-preS1, then an indirect ELISA was done. Apparent affinity was determined as the reciprocal of the antigen concentration required to inhibit 50% maximal binding in the competitive ELISA.

Hong, H. J., Ryu, C. J., Hur, H., Kim, S., Oh, H. K., Oh, M. S., & Park, S. Y. (2004). In vivo neutralization of hepatitis B virus infection by an anti-preS1 humanized antibody in chimpanzees. Virology, 318(1), 134-141.