Combining phage display with SMRTbell next-generation sequencing for the rapid discovery of functional scFv fragments by Francesco Nannini, Lenart Senicar, Farhaan Parekh, Khai Kong, Alexander Kinna, Reyisa Bughda, James Sillibourne, Xihao Hu, Biao Ma, Yuchen Bai, Mathieu Ferrari, Martin Pule, Shimobi Onuoha.
Phage display technology in combination with next-generation sequencing (NGS) currently is a state-of-the-art method for the enrichment and isolation of monoclonal antibodies from diverse libraries. However, current NGS methods employed phage libraries are limited by short contiguous read lengths associated second-generation platforms. Consequently, identification antibody sequences has conventionally been restricted to individual domains or analysis single domain binding moieties such as camelid VHH cartilaginous fish IgNAR antibodies. In this study, we report application third-generation address limitation. We used molecule real time (SMRT) coupled hairpin adaptor loop ligation facilitate accurate interrogation full-length single-chain Fv (scFv) Our facilitated rapid testing scFv enriched within days following panning. Two against CD160 CD123 were panned monitored NGS. Analysis data sets led several functional that not identified conventional panning screening strategies. approach, which combines selection immune fragments, an easy discover antibodies, range affinities biophysical characteristics.
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