期刊
JOURNAL OF MEDICAL MICROBIOLOGY
卷 67, 期 6, 页码 820-827出版社
MICROBIOLOGY SOC
DOI: 10.1099/jmm.0.000744
关键词
single-chain variable fragment (scFv); ribosome display; Neisseria meningitidis; recombinant protein; antibody; Escherichia coli
类别
资金
- Presidency of the Islamic Republic of Iran National Elites Foundation [91002087]
Purpose. Neisseria meningitidis is the leading global cause of meningitis and sepsis. Detection, followed by identification, of bacterial pathogens is important in medicine and public health. In the present study, we used the ribosome display technique to select single-chain variable fragments (scFv) that are specific to the surface-exposed fHbp antigen of N. meningitidis. Methodology. The recombinant fHbp protein was used as the antigen for the immunization of BALB/c mice. Anti-fHbp VH/k chain ribosome display libraries were assembled by joining VH and k into the VH/k chain with a specially constructed linker by PCR overlap extension. The scFv library was panned against the recombinant fHbp protein by using a single round of the ribosome display method via a rabbit reticulocyte lysate system. Results/Key findings. The selected anti-fHbp antibody exhibited high affinity and specificity in the enzyme-linked immunosorbent assay (ELISA) and the whole bacterial cell enzyme-linked immunosorbent assay (Bact-ELISA). Conclusion. The affinity of the selected scFv was similar to 8.65 x 10(9) M-1. The isolated scFv can provide the basis for developing a diagnostic kit.
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