Figure 2.

Schematic overview of the strategy used for construction of antigen specific phage library. The peripheral blood mononuclear cells (PBMCs) were isolated from an antiretroviral drug naïve patient (#254) whose plasma exhibited neutralizing activity against a panel of viruses and also displayed cross clade reactive anti-V3 antibody binding potential. PBMCs were subjected to EBV transformation in 96 well culture plate, wells showing high level of anti-V3 antibodies were selected and expanded to 24 well stage and then to six well stage . The wells showing high titre of anti-V3 Abs were pooled together and cultured in T25 flask. Total RNA was isolated from these antigen specific enriched B cells and cDNA was synthesised. Heavy and light chains were amplified and scFvs were constructed and cloned into a pCANTAB-5E phagemid vector. scFv phage library of 7000 clones was constructed.