Fig. 4.
Indirectly screening of bacteriophage libraries. (A) The enrichment of library for phage display. (B) Monoclonal phage SPR of the selected anti-BoNT phages binding to AHCC particles. Anti-M13 antibody was immobilized on the CM5 chip to capture phage antibody particles in the supernatant, and then the dissociation and association to bind 40 nM AHCC was determined. Eight candidate clones with high Ru values were selected for subsequent validation. (C) Cell supernatants of clones were obtained by transient expression of candidate antibodies fused with Fc in 293T cells. The Protein A chip capture antibodies in the supernatant, respectively, and then the dissociation and association rates to bind to AHCC were determined. (D) SPR analyses the binding of AHCC to HM. The Protein A chip capture HM, AHCC: 10 nM, 5 nM, 2.5 nM, 1.25 nM, 0.625 nM and 0.3125 nM.