Fig. 2.

Isolation and efficacy of CCK-BR scFv antibodies. A. Schematic of stalled ARM complex and position of primers used for RT-PCR recovery in the 1st, 2nd, and 3rd cycles of ribosome display. B. Analysis of RT-PCR recovery of VH/K cDNA from CCK-B receptor immunized spleen in the 1st, 2nd, and 3rd cycles. C. Western blot of seven purified unique CCK-BR scFvs generated by the cell-free ribosome display platform. D. Plot of ELISA data showing binding affinity of increasing concentrations of the three scFv antibodies with the best binding. E. Binding specificity and cross-reactivity of 3 CCK-BR scFvs to CCK-BR, but not CCK-AR or P2X4R. F. Immunoblot of recombinant cholecystokinin-B receptor (CCK-BR) antigen by soluble single-chain variable fragment (scFv) antibody fragments. a. Purified scFvs. b. Unrelated scFv as a negative control. c. A goat anti-CCK-BR polyclonal antibody as a positive control.