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V. Tiwari, C. Clement, D. Shukla; Isolation of Novel Peptides Against Cell Surface Heparan Sulfate . Invest. Ophthalmol. Vis. Sci. 2006;47(13):3078.
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© ARVO (1962-2015); The Authors (2016-present)
Cell surface heparan sulfate (HS) including the modified from of HS is crucial for many biological processes including the pathogenesis of herpes simplex virus–1 (HSV–1) in the cornea. The goals of this study were to isolate 12–mer peptides against cell surface HS to study structural–functional aspects of HS.
A phage display library (12 mer, New England Biolab) was pre–screened against a blocking reagent (BSA). The non binding phages recovered from the supernatant were then used to screen commercially available HS. During 3 panning cycles against the HS target, the tween–20 concentration was increased from0.2% to 0.5% to raise to stringency of washing condition and produce higher affinity binding phages. The bound phages were recovered, plaque purified and tittered. This was followed by DNA sequencing of the phage–encoded peptides. The isolated phages were tested for their effect on viral infectivity to various cell–types.
The amino acid sequences of isolated peptides against HS revealed some uniquely placed positively charged amino acids indicating spatial requirements for binding to HS. Interestingly, some of the isolated peptides showed promising results by interfering with HSV–1 entry.
Using the phage display technique we identified novel peptides against cell surface HS that could be used to block HSV–1 entry.
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