Fig. 2: CD151 in situ Ab selection process. | Communications Biology

Fig. 2: CD151 in situ Ab selection process.

From: CellectSeq: In silico discovery of antibodies targeting integral membrane proteins combining in situ selections and next-generation sequencing

Fig. 2

Circles represent live HEK293T cell lines of either overexpressing CD151 (HEK293T-CD151+; denoted by “+” sign), or CD151 knockdown cells (HEK293T-CD151−; denoted by a “−” sign). For round 1, the Fab-phage (naïve library F) were incubated with HEK293T-CD151+ cells, then eluted and amplified for the next round. For rounds 2 and 3, the Fab-phage were first incubated with HEK293T-CD151− cells, then transferred and incubated with HEK293T-CD151+ cells. For round 4, amplified round 3 Fab-phage were independently incubated with HEK293T-CD151− or HEK293T-CD151+ cells. The dashed lines with arrows indicate that eluted phage from the preceding round were amplified through E. coli prior to incubation with cells in the succeeding round. The solid lines with arrows indicate that unbound phage from the preceding cell line were transferred directly to the succeeding cell line. In round 4, phages from Fab-phage pools HEK293T-CD151+ (Positive pool) and HEK293T-CD151− (Negative pool) were amplified and used as DNA template for NGS analysis.

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