pH-dependent antibody engineering via yeast surface display
A worked technical case study from a real client campaign. Six rounds of FACS selection across a 640-clone library converged on a small set of hotspot substitutions that switch antibody binding between physiological and endosomal pH.
Download the case study
14 pages, PDF. Enter your details for instant access.
A complete walkthrough of the campaign, from library design to ranked candidates
Library design and quality control
640 targeted variants, NGS-confirmed coverage and uniformity before selection begins. Diversity rationale and the trade-offs that informed library size.
FACS panel and gating strategy
Two-color labeling for surface expression and antigen binding, paired pH-7.4 and pH-5.5 sort arms, gating thresholds tuned per cycle to maintain enrichment without losing diversity.
Enrichment-score methodology
How per-cycle read counts are converted into a quantitative enrichment metric. Why enrichment scores rank variants more reliably than raw read counts at any single cycle.
Convergent hotspot analysis
Two residues (H-1 and H-2) recurred across independent enriched lineages. Convergence is the strongest signal that a position genuinely controls pH-dependent binding rather than reflecting library bias.
Cumulative enrichment ranking
Top candidates ranked by cumulative enrichment across all six cycles. Why a multi-cycle composite outperforms picking winners from any single sort.
Decision framework for pH-dependent campaigns
When pH-dependent binding is worth engineering for (FcRn-mediated recycling, tumor-microenvironment selectivity, ADC linker stability) and how to scope library size and sort cycle count for your target.
Run a yeast display campaign for your target
Tell us about your binder, your target, and the property you want to engineer. We will scope a campaign and return a timeline within 48 hours.
Start a project →