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[Construction of a human single-chain fragment antibody against severe fever with thrombocytopenia syndrome virus].

Objective To construct a human phage display library against severe fever with thrombocytopenia syndrome (SFTS) virus. Methods The total RNA was isolated from peripheral blood lymphocytes of 8 patients with SFTS and cDNA was amplified. The genes of light-chain (Vκ and Vλ) and heavy-chain (VH) were amplified by PCR. The scFv gene was linked by overlap-PCR and cloned into the vector pComb3XSS, and then transformed into XL1-Blue cells for the phage antibody library construction. After detecting the recombinant rate and the library repertoire, we screened the antibodies against SFTS virus by biopanning with immobilized virus antigen. Results The volume of constructed phage antibody library was 2.8×107 . The results of sequencing showed that the sequence of the contained single-chain variable fragment (scFv) was different. After three rounds of panning, phage antibodies were specifically enriched. A total of 21 clones were positive against SFTS virus by phage-ELISA. Conclusion A high-capacity and diverse human scFv against SFTS virus has been constructed successfully.

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