Peptides on phage: a vast library of peptides for identifying ligands. Export

@article{citeulike:5892868, abstract = {We have constructed a vast library of peptides for finding compounds that bind to antibodies and other receptors. Millions of different hexapeptides were expressed at the N terminus of the adsorption protein (pIII) of fd phage. The vector fAFF1, derived from the tetracycline resistance-transducing vector fd-tet, allows cloning of oligonucleotides in a variety of locations in the 5' region of gene III. A library of 3 x 10(8) recombinants was generated by cloning randomly synthesized oligonucleotides. The library was screened for high-avidity binding to a monoclonal antibody (3-E7) that is specific for the N terminus of beta-endorphin (Tyr-Gly-Gly-Phe). Fifty-one clones selected by three rounds of the affinity purification technique called panning were sequenced and found to differ from previously known ligands for this antibody. The striking finding is that all 51 contained tyrosine as the N-terminal residue and that 48 contained glycine as the second residue. The binding affinities of six chemically synthesized hexapeptides from this set range from 0.35 microM (Tyr-Gly-Phe-Trp-Gly-Met) to 8.3 microM (Tyr-Ala-Gly-Phe-Ala-Gln), compared with 7.1 nM for a known high-affinity ligand (Tyr-Gly-Gly-Phe-Leu). These results show that ligands can be identified with no prior information concerning antibody specificity. Peptide libraries are also likely to be useful in finding ligands that bind to other classes of receptors and in discovering pharmacologic agents.}, author = {Cwirla, S. E. and Peters, E. A. and Barrett, R. W. and Dower, W. J.}, citeulike-article-id = {5892868}, citeulike-linkout-0 = {http://view.ncbi.nlm.nih.gov/pubmed/2201029}, citeulike-linkout-1 = {http://www.hubmed.org/display.cgi?uids=2201029}, issn = {0027-8424}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, keywords = {phage\_display, protocol, thesis}, month = {August}, number = {16}, pages = {6378--6382}, posted-at = {2009-10-05 21:06:45}, priority = {2}, title = {Peptides on phage: a vast library of peptides for identifying ligands.}, url = {http://view.ncbi.nlm.nih.gov/pubmed/2201029}, volume = {87}, year = {1990} }

TY - JOUR ID - citeulike:5892868 L3 - citeulike-article-id:5892868 N2 - We have constructed a vast library of peptides for finding compounds that bind to antibodies and other receptors. Millions of different hexapeptides were expressed at the N terminus of the adsorption protein (pIII) of fd phage. The vector fAFF1, derived from the tetracycline resistance-transducing vector fd-tet, allows cloning of oligonucleotides in a variety of locations in the 5' region of gene III. A library of 3 x 10(8) recombinants was generated by cloning randomly synthesized oligonucleotides. The library was screened for high-avidity binding to a monoclonal antibody (3-E7) that is specific for the N terminus of beta-endorphin (Tyr-Gly-Gly-Phe). Fifty-one clones selected by three rounds of the affinity purification technique called panning were sequenced and found to differ from previously known ligands for this antibody. The striking finding is that all 51 contained tyrosine as the N-terminal residue and that 48 contained glycine as the second residue. The binding affinities of six chemically synthesized hexapeptides from this set range from 0.35 microM (Tyr-Gly-Phe-Trp-Gly-Met) to 8.3 microM (Tyr-Ala-Gly-Phe-Ala-Gln), compared with 7.1 nM for a known high-affinity ligand (Tyr-Gly-Gly-Phe-Leu). These results show that ligands can be identified with no prior information concerning antibody specificity. Peptide libraries are also likely to be useful in finding ligands that bind to other classes of receptors and in discovering pharmacologic agents. IS - 16 JF - Proceedings of the National Academy of Sciences of the United States of America SN - 0027-8424 EP - 6382 TI - Peptides on phage: a vast library of peptides for identifying ligands. VL - 87 SP - 6378 KW - phage_display KW - protocol KW - thesis AU - Cwirla, SE AU - Peters, EA AU - Barrett, RW AU - Dower, WJ PY - 1990/08// UR - http://view.ncbi.nlm.nih.gov/pubmed/2201029 ER -