PMID: 7536850Apr 14, 1995Paper

Comprehensive epitope analysis of monoclonal anti-proenkephalin antibodies using phage display libraries and synthetic peptides: revelation of antibody fine specificities caused by somatic mutations in the variable region genes

Journal of Molecular Biology
V BöttgerB A Spruce

Abstract

Filamentous phage libraries, displaying 6, 12 or 20 amino acid residue peptides at the N terminus of coat protein pIII were used to define and localize the epitopes of 15 monoclonal antibodies raised against human proenkephalin, a neuropeptide precursor. Eight monoclonal antibodies (PE14 to PE19, PE23 and PE25), which inhibit each other's binding to proenkephalin, recognized phage clones selected by PE14, PE15, PE19, PE23 and PE25. With the peptide sequences DLL(X)(X)LL (12mer library) and DLL(X)(X)L (6mer library) shared by most of the phage clones it was possible to define the putative antibody epitope 155DLLKELL161 on human proenkephalin. For five antibodies (PE13, PE20 to PE22 and PE24) belonging to another inhibition group, a common consensus motif G(X)D(X)E(X)(X)V(X)(X)R could be defined with help of a 20mer library. The corresponding minimum epitope sequence has been found to be 175GSDNEEEVSKR185. Antibody PE1, raised in a separate fusion, was able to select phage clones from a 12mer and 20mer library, revealing that the sequence 187GGFMRG192 is probably the antibody epitope. The assumed localization of the epitopes was confirmed by screening a set of overlapping synthetic peptides, covering the region of human proenkeph...Continue Reading

Citations

Jul 1, 1997·Protein Science : a Publication of the Protein Society·T Pacholczyk, K J Sweadner
Sep 1, 1995·The Journal of Cell Biology·A Böttger, B A Spruce
Mar 17, 1999·Proceedings of the National Academy of Sciences of the United States of America·S P WilsonJ C Glorioso
Feb 28, 2014·Gene Therapy·A Z TzabazisD C Yeomans
Mar 3, 2010·BMC Genomics·Jagadeesh N MavinahalliChandra Verma
Jun 27, 1997·Journal of Molecular Biology·A BöttgerD P Lane
Feb 24, 2010·Parasitology Research·Aijiang GuoXuenong Luo
Feb 2, 1999·Oncogene·V BöttgerD P Lane
Jul 9, 1999·Journal of Molecular Recognition : JMR·V BöttgerB Micheel
Aug 26, 1998·The Journal of Clinical Endocrinology and Metabolism·S ChenJ Furmaniak
Mar 10, 2001·The Journal of Immunology : Official Journal of the American Association of Immunologists·M M Souto-CarneiroV Krenn
Feb 28, 2001·Journal of Immunological Methods·L ChoulierD Altschuh
Apr 1, 1997·Chemical Reviews·George P. Smith, Valery A. Petrenko

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