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PEDS Advance Access published online on November 6, 2008
Protein Engineering Design and Selection, doi:10.1093/protein/gzn060
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© The Author 2008. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oxfordjournals.org

Diversity and censoring of landscape phage libraries

G.A. Kuzmicheva1, P.K. Jayanna, I.B. Sorokulova2 and V.A. Petrenko3

Department of Pathobiology, Auburn University, t252 Greene Hall, College of Veterinary Medicine, Auburn, AL 36849, USA

3 To whom correspondence should be addressed. E-mail: petreva{at}vetmed.auburn.edu

Libraries of random peptides displayed on the surface of filamentous phages are a valuable source for biospecific ligands. However, their successful use can be hindered by a disproportionate representation of different phage clones and fluctuation of their composition that arises during phage reproduction, which have potential to affect efficiency of selection of clones with an optimal binding. Therefore, there is a need to develop phage display libraries with extended and varied repertoires of displayed peptides. In this work, we compared the complexity, evolution and representation of two phage display libraries displaying foreign octamers and nonamers in 4000 copies as the N-terminal part of the major coat protein pVIII of phage fd–tet (landscape libraries). They were obtained by replacement of amino acids 2–4 and 2–5 of pVIII with random octa- and nonamers, respectively. Statistical analysis of the libraries revealed their dramatic censoring and evolution during amplification. Further, a survey of both libraries for clones that bind common selectors revealed the presence of different non-overlapping families of target-specific clones in each library justifying the concept that different landscape libraries cover different areas of a sequence space.

Keywords: landscape phage/phage display/phage library/Salmonella typhimurium/streptavidin

Received June 13, 2008; revised September 30, 2008; accepted October 7, 2008.

1 Present address: Vegrandis LLC, 700 W. Research Blvd, Fayetteville, AR 72701, USA

2 Present address: Department of Anatomy, Physiology and Pharmacology, Auburn University, 109 Greene Hall, College of Veterinary Medicine, Auburn, AL 36849, USA

Abbreviations: AU, absorbance units; OD, optical density; CFU, colony-forming units; vir-virions; RT, room temperature; SA, streptavidin; PEG, polyethyleneglycol; NPP, p-nitrophenyl phosphate; AP-SA, alkaline phosphatase-conjugated streptavidin


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Home page
 Protein Eng Des SelHome page
G.A. Kuzmicheva, P.K. Jayanna, A.M. Eroshkin, M.A. Grishina, E.S. Pereyaslavskaya, V.A. Potemkin, and V.A. Petrenko
Mutations in fd phage major coat protein modulate affinity of the displayed peptide
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