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Giang KA, Nygren PÅ, Nilvebrant J
Methods in molecular biology (Clifton, N.J.) 2702 (-) 373-392 [2023-09-08; online 2023-09-08]
Herein, we describe a general protocol for the selection of target-binding affinity protein molecules from a phagemid-encoded library. The protocol is based on our experience with phage display selections of non-immunoglobulin affibody affinity proteins but can in principle be applied to perform biopanning experiments from any phage-displayed affinity protein library available in a similar phagemid vector. The procedure begins with an amplification of the library from frozen bacterial glycerol stocks via cultivation and helper phage superinfection, followed by a step-by-step instruction of target protein preparation, selection cycles, and post-selection analyses. The described procedures in this standard protocol are relatively conservative and rely on ordinary reagents and equipment available in most molecular biology laboratories.
PubMed 37679630
DOI 10.1007/978-1-0716-3381-6_19
Crossref 10.1007/978-1-0716-3381-6_19