Selection of Apoptotic Cell Specific Human Antibodies from Adult Bone Marrow
Figure 1
Phage display selection of human antibodies binding to PC and ACs.
The first three rounds of panning used PC conjugated to BSA in solution, while cycle four and five amplified binding to whole apoptotic Jurkat cells. The amplification of binding phage in the selection is illustrated in (A) showing the titers of bound phage eluted in each selection cycle, and (B) showing an increase in the ratio of eluted phage (phage out) of the number of phage particles used going into each round (phage in) presented as % on a logarithmic scale and (C) showing the increase of PC-binding in ELISA of the starting phage libraries in each selection cycle at 1∶5 dilution and detection with anti-M13 phage. Randomly selected individually clones were cultured from selection round five and screened for antibody expression and PC-binding by ELISA using anti-Fab detection. D. Screening for PC-binding (black) compared to binding to the control antigen MDA-BSA (white). E. Screening for human Fab (black) and kappa light chain (white) expression. The data in both D and E were sorted based on PC reactivity from high to low. Bacterial supernatant without Fab-expressing phagemid was used as negative control and human IgG1 (κ) as positive control.