TY - JOUR
T1 - Dual surface selection methodology for the identification of thrombin binding epitopes from hotspot biased phage-display libraries
AU - Rajagopal, Srivats
AU - Meza-Romero, Roberto
AU - Ghosh, Indraneel
N1 - Funding Information:
This work was supported by the Research Corporation (Research Innovation Award), ACS (PRF TypeG) and the Leukemia and Lymphoma Society of America. We thank Min Zhou and Scott Meyer for helpful advice.
PY - 2004/3/22
Y1 - 2004/3/22
N2 - 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.
AB - 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.
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U2 - 10.1016/j.bmcl.2003.09.098
DO - 10.1016/j.bmcl.2003.09.098
M3 - Article
C2 - 15006368
AN - SCOPUS:1542315257
SN - 0960-894X
VL - 14
SP - 1389
EP - 1393
JO - Bioorganic and Medicinal Chemistry Letters
JF - Bioorganic and Medicinal Chemistry Letters
IS - 6
ER -