FIG 7.
Repertoire analysis of somatic variants of IGHV1-69-encoded mAbs. Deep sequence analysis of antibody variable genes expressed in donor PBMCs revealed somatic variants for the heavy chains of STAU-399 and -229. These somatic variant heavy chain antibodies were expressed recombinantly with the original light chain and purified. (a and b) Neighbor-joining tree analysis of mAb clonal variants by Geneious. The germ line gene sequences of IGHV1-69*01 or IGHV1-69-2*01 were used as the outgroup sequences. (c and d) Each variant mAb was tested for binding to IsdB, and the half-maximal effective concentration (EC50) for binding was determined. Biolayer interferometry was used to determine the on and off rates of each variant antibody for binding to NEAT2, and the calculated KD is graphed. These experiments were performed three times independently.