Fig. 2.
Schematic illustrating affinity purification technique. Clarified pooled antisera was diluted in binding buffer. The diluted serum was applied to the five depletion columns – GST, EG954–74, EG9554–109, EG9592–156 and E. coli lysate. The flow through was reapplied to the depletion columns three times. Washed columns were eluted of antibodies. Anti-GST antibodies were collected. The flow through from the five depletion columns was applied to an EG95–GST column. The flow through from this column was reapplied three times. Washed EG95–GST column was eluted of antibodies and the antibodies against conformational epitopes on EG95 were collected (anti-cEG95).