Screening and identification of HIV-1 gp41 core structure epitope from random phage display peptide library
Bei Liu
Abstract
Bei Liu
Abstract
Objective:To identify and characterize the epitopes on core structure of HIV 1 gp41.Methods:A random phage displayed dodecapeptide library was screened with a conformation specific monoclonal antibody NC 1 specifically against the core structure of HIV 1 gp41.The positive clones were identified by sandwich ELISA,soluble NC 1 blocking assay and competitive inhibition assay.Results:After three rounds of screening,10 of 24 phage clones were identified as positive clones which can bind to NC 1.Amino acid sequences deduced from DNA sequences showed five different sequences:HDVHHRWVYLLS?ITVNEWLYTSEQ?HGRSHGMFKPKR?MGPIARPHWHLN?DMYRSPRPKPDT.The binding between phage clones(displayed HDVHHRWVYLLS,VNEWLYTSEQ and MGPIARPHWHLN, respectively)and NC 1 could be inhibited by N36 C34 complex.Soluble NC 1 could block the binding between phage clones and NC 1.Conclusion:The results indicate that HDVHHRWVYLLS,VNEWLYTSEQ and MGPIARPHWHLN are the mimotopes which could mimic the core structrue epitopes of six helix bundle of HIV 1 gp41.
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Objective:To identify and characterize the epitopes on core structure of HIV 1 gp41.Methods:A random phage displayed dodecapeptide library was screened with a conformation specific monoclonal antibody NC 1 specifically against the core structure of HIV 1 gp41.The positive clones were identified by sandwich ELISA,soluble NC 1 blocking assay and competitive inhibition assay.Results:After three rounds of screening,10 of 24 phage clones were identified as positive clones which can bind to NC 1.Amino acid sequences deduced from DNA sequences showed five different sequences:HDVHHRWVYLLS?ITVNEWLYTSEQ?HGRSHGMFKPKR?MGPIARPHWHLN?DMYRSPRPKPDT.The binding between phage clones(displayed HDVHHRWVYLLS,VNEWLYTSEQ and MGPIARPHWHLN, respectively)and NC 1 could be inhibited by N36 C34 complex.Soluble NC 1 could block the binding between phage clones and NC 1.Conclusion:The results indicate that HDVHHRWVYLLS,VNEWLYTSEQ and MGPIARPHWHLN are the mimotopes which could mimic the core structrue epitopes of six helix bundle of HIV 1 gp41.
Key concepts: Epitope, Gp41, Phage display, Monoclonal antibody, Mimotope, Molecular biology, Peptide library, Biology