Searching sequence space for high-affinity binding peptides using ribosome display.

Thorsten Lamla; Volker A Erdmann

doi:10.1016/s0022-2836(03)00432-7

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Searching sequence space for high-affinity binding peptides using ribosome display.

Thorsten Lamla·Volker A Erdmann

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Abstract

We present the construction of a synthetic library based on the scaffold of bovine heart fatty acid-binding protein (FABP) with 1.1x10(14) independent members. Ribosome display was applied to select streptavidin-binding peptides in vitro from 2x10(13) molecules of the library each encoding FABP with 15 contiguous random amino acid residues at its N terminus. The selection yielded several different binding peptides. The best binder possessed a dissociation constant as low as 4nM and, in contrast to the previously isolated peptides, contained no HPQ motif. A substitution analysis enabled shortening of the 15-mer peptide and revealed a 9-mer variant with a dissociation constant of 17nM, which is a 1000-fold increase of affinity compared to the already known peptides of this size. This high-affinity binding peptide in combination with the whole set of streptavidin conjugates should be an extremely useful tool for the detection and purification of recombinant proteins.

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Animals Binding Sites Carrier Proteins Cattle DNA Primers Fatty Acid-Binding Proteins Heart In Vitro Techniques Neoplasm Proteins Oligopeptides Peptide Library Peptides Polymerase Chain Reaction RNA, Messenger Ribosomes Streptavidin Transcription, Genetic Journal Article Research Support, Non-U.S. Gov't

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Thorsten Lamla, Volker A Erdmann. 2003-05-30. Searching sequence space for high-affinity binding peptides using ribosome display.. https://doi.org/10.1016/s0022-2836(03)00432-7

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