Fig. 5: Binding analysis of lead cyclonal antibodies selected from combinatorial libraries.
a Comparison of CDRs for murine anti-HAG cyclonal and human 20.1 and 75.1 cyclonals. Asterisk (*) indicates residue shared by all three clones; plus sign (+) indicates residue shared by two clones. b ELISA analysis of library-selected cyclonal antibodies derived from genetic selection with HAG as target antigen. Binding activity and specificity of the lead human cyclonal antibodies, 20.2 and 75.1, as well as the murine anti-HAG and anti-Gcn4 cyclonals were evaluated by ELISA using purified GST-HAG or BSA as immobilized antigen. Absorbance was measured at 450 nm for each cyclonal and the resulting measurements were analyzed using GraphPad Prism 9 to determine binding affinity. Data are the average of three biological replicates ± SD. c Biolayer interferometry (BLI) kinetic assays were performed to measure kinetic binding constants (ka, kd) and equilibrium binding constants (KD) for the interaction between the 20.2 and 75.1 cyclonals and HAG peptide. Biotinylated GST-HAG was immobilized on streptavidin (SA)-coated sensors and subsequently used to bind protein A-purified cyclonals in solution. Response data are representative of replicate BLI experiments (n = 2). Source data are provided as a Source Data file.
