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- W2365694555 abstract "Objective To find specific polypeptides which bind RSV with high affinity and inhibit replication of RSV, we screened ligands on intact RSV virion by display libraries of phage. Methods Human RSV was harvested in Hep 2 cells. The PEG concentrated virus was layered over sucrose density gradients (30%, 45%, 60%), followed by ultrafiltration. The purified RSV was biotinylated in vitro with NSH LC biotin and then to react with random peptide library displaying 7 amino acids fused on protein Ⅲ of M13 phage. The selected peptides for target binding were assayed by ELISA after 3 rounds of biopanning and measured by 50 percent tissue culture infection dose (TCID 50 ). The positive clones with high affinity were used for automated sequencing with dye labeled dideoxynucleotides, and the amino acid sequence of polypeptide displayed on phage was deduced. Results The enrichment was shown by ELISA after 3 rounds of biopanning and 13 positive clones bound to coat protein of RSV with high affinity. Three positive clones were identified to inhibit the replication of RSV in Hep2 cells in vitro that decreased RSV TCID 50 from 10 -8.1 SFU/ml to 10 -4.1 , 10 -4.3 , 10 -3.8 SFU/ml respectively. Sequencing of the genes encoding these peptides in 13 positive clones showed absence of any conserved motifs, although most peptides had high proportion of argenines. Conclusion Positive clones against RSV can be selected from phage display peptide library and so provide a potential tool for highly sensitive diagnostic kits and novel antiviral agents." @default.
- W2365694555 created "2016-06-24" @default.
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- W2365694555 date "2002-01-01" @default.
- W2365694555 modified "2023-09-24" @default.
- W2365694555 title "Anti-RSV polypeptides selected from a phage displayed random peptide library" @default.
- W2365694555 hasPublicationYear "2002" @default.
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