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Isolation of Human Single Chain Antibody against Fusion Protein of Human Respiratory Syncytial Virus |
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Abstract Objective:To obtain the human single chain antibody (scFv) against fusion protein of Respiratory Syncytial Virus. Methods:A na?ve library of phage display human scFv, genes of which were derived from peripheral blood lymphocytes (PBLs) of 10 healthy donors, was constructed and panned by F of RSV to select specific anti-RSV F scFv. After 5 rounds of panning, the activity of individual clone was examined by ELISA. The genes of positive clones were sequenced. Soluble scFv was prepared through infecting E.coli HB2151 with one of the positivephage clones. Results:After the panning, scFvs from 18 clones can bind F specifically by ELISA. DNA sequence analysis of the clone E4 with the highest OD indicates that the variable region gene of the clone is highly homologous with human immunoglobulin gene family. The expression of scFv E4 has been confirmed by Western and Dot blots. Conclusion:The specific human anti-RSV F scFv has been prepared successfully by exploiting phage antibody library.
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Received: 15 May 2009
Published: 10 May 2010
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