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• W2358964798 abstract "Objective To screen the mimotopes of CFP-10/ESAT-6 (CE) fusion protein from mycobacterium tuberculosis using phage-displayed random peptide libraries. Methods The monoclonal antibody against the CE fusion protein was used to screen the binding peptide from the Ph.D.-7 peptide library. After 3 rounds of screening, 18 clones were randomly selected for DNA sequencing. The selected clones were tested by sandwich enzyme linked immunosorbent ELISA and competition ELISA. By indirect ELISA method, antibodies against single-chain phage positive clones and CE fusion protein were detected in serum samples from 20 patients with tuberculosis and 10 healthy people who had been vaccinated with BCG. Results After three rounds of biopanning, remarkable enrichment of the phage that can specifically bind with target molecules were observed.By DNA screening, 9 sequences were obtained, in which the sequences of single-chain phages 1, 7, 11, 12, 15, and 16 were exactly the same, and the conservative sequence Trp-Asp-Ala-Thr (WDAT), which was identical with the sequence from the 58th to 61st amino acids in ESAT-6, was found within the amino acid sequences of phages 5, 6, and 18. Nine phages with different sequences were detected using sandwich enzyme linked immunosorbent ELISA and competition ELISA, and 7 of them were found to be positive clones with immunoactivity (the S/N value of the phages 1, 5, 6, 10,13, 14, and 18 were 9.2, 9.7, 9.4, 8.9, 9.6, 9.9, and 9.0, respectively). Indirect ELISA using the positive phage 5 that containing WDAT sequence or CE fusion protein showed that the absorbance value of both the serum samples detected by using phage 5 were higher than that by CE fusion protein (0.931 ± 0.298 vs 0.317 ± 0.157, 0.496 ± 0.073 vs 0.118 ± 0.026, all P 0.05). The detection rate of the serum samples from patients with tuberculosis by using phage 5 (95%, 19/20) that was significantly higher than that by using CE fusion protein (60%, 12/20), but the detection rate of the serum samples from healthy people was 9/10, which was lower than that by using CE fusion protein (10/10). Conclustions By using phage-displayed random peptide libraries, we screened out 7 mimotopes of CE fusion protein, and a liner B cell epitope positioned at the sequence of the 58th to 61st amino acids at ESAT-6. Basing on the results, the sensitivity of the ELISA test can be increased and detection reagent for anti-tuberculosis antibodies may be developed." @default.
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• W2358964798 date "2008-01-01" @default.
• W2358964798 modified "2023-09-26" @default.
• W2358964798 title "Screening of mimotopes of CFP-10/ESAT-6 fusion protein from mycobacterium tuberculosis" @default.
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