Dual surface selection methodology for the identification of thrombin binding epitopes from hotspot biased phage-display libraries

Srivats Rajagopal, Roberto Meza-Romero, Indraneel Ghosh

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Protein libraries biased towards amino-acid residues found at so-called 'hotspots' were incorporated into the beta-sheet region of the thermostable variant (HTB1) of the B1 domain of the immunoglobulin (IgG) binding protein G and expressed as gene 3 fusions on M13 bacteriophage. The HTB1 library (2.2×109) variants with a minimal 12 amino acid basis set were selected for binding IgG, to ensure structural conservation, and subsequently to thrombin to evolve a thrombin-binding function. We believe that this dual surface selection strategy will have great utility in evolving new bi-functional proteins without compromising structure. Furthermore the discrete beta-sheet epitopes identified by our methodology will lend itself to small-molecule mimicry of beta-sheets.

Original languageEnglish (US)
Pages (from-to)1389-1393
Number of pages5
JournalBioorganic and Medicinal Chemistry Letters
Volume14
Issue number6
DOIs
StatePublished - Mar 22 2004

Fingerprint

Bacteriophages
Thrombin
Libraries
Epitopes
Immunoglobulin G
Display devices
Amino Acids
Bacteriophage M13
Immunoglobulins
Conservation
Carrier Proteins
Proteins
Fusion reactions
Genes
Gene Fusion
Molecules
beta-Strand Protein Conformation

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Organic Chemistry
  • Drug Discovery
  • Pharmaceutical Science

Cite this

Dual surface selection methodology for the identification of thrombin binding epitopes from hotspot biased phage-display libraries. / Rajagopal, Srivats; Meza-Romero, Roberto; Ghosh, Indraneel.

In: Bioorganic and Medicinal Chemistry Letters, Vol. 14, No. 6, 22.03.2004, p. 1389-1393.

Research output: Contribution to journalArticle

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