Figure 1

The outline of the subtractive panning campaign. PEPI-AC, AC and PTM were immobilized overnight on microtiter plate well. The library phage was pre-incubated with PTM in PTM coated well and the process was repeated twice (well 1–3). Subsequently, the pre-incubated library phage was transferred to AC coated well to capture and eliminate AC binders from the library phage pool. The process was repeated twice (well 4–6). Lastly, the library phage was transferred to PEPI-AC coated well to isolate specific binders. Bound phage was eluted and recovered by infecting TG1 and plate out on ampicillin supplemented agar plate. The colonies were scraped and culture in 10 mL 2 × YT supplemented with ampicillin and glucose. The culture was infected with M13KO7 helper phage when OD_600nm 0.5 was reached. After overnight packaging, the culture was pellet down and phages secreted in the medium were precipitated with PEG/NaCl. The recovered phages were resuspended with PBS and use for polyclonal and monoclonal ELISA.