Quantitative synthesis of genetically encoded glycopeptide libraries displayed on M13 phage.
Phage display is a powerful technology that enables the discovery of peptide ligands for many targets. Chemical modification of phage libraries have allowed the identification of ligands with properties not encountered in natural polypeptides. In this report, we demonstrated the synthesis of 2 × 10(8) genetically encoded glycopeptides from a commercially available phage-displayed peptide library (Ph.D.-7) in a two-step, one-pot reaction in <1.5 h. Unlike previous reports, we bypassed genetic engineering of phage. The glycan moiety was introduced via an oxime ligation following oxidation of an N-terminal Ser/Thr; these residues are present in the peptide libraries at 20-30% abundance. The construction of libraries was facilitated by simple characterization, which directly assessed the yield and regioselectivity of chemical reactions performed on phage. This quantification method also allowed facile yield determination of reactions in 10(9) distinct molecules. We envision that the methodology described herein will find broad application in the synthesis of custom chemically modified phage libraries.
Alberta Glycomics Centre and Department of Chemistry, University of Alberta, Edmonton, AB T6G 2G2, Canada.
Simon
S
Ng
Mohammad R
MR
Jafari
Wadim L
WL
Matochko
Ratmir
R
Derda
D017104:Bacteriophage M13 / Q000737:chemistry* / Q000235:genetics; D006020:Glycopeptides / Q000138:chemical synthesis* / Q000235:genetics; D010084:Oxidation-Reduction; D010091:Oximes / Q000737:chemistry*; D019151:Peptide Library; D011134:Polysaccharides / Q000737:chemistry
D006020:Glycopeptides; D010091:Oximes; D019151:Peptide Library; D011134:Polysaccharides