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Preparation of a Single-Chain Antibody against Nucleocapsid Protein of Porcine Deltacoronavirus by Phage Display Technology.
Zhang, Yixuan; Song, Yue; Ren, Haojie; Zeng, Quan; Yuan, Yixin; Xia, Lu; Wei, Zhanyong.
  • Zhang Y; College of Veterinary Medicine, Henan Agricultural University, Zhengzhou 450046, China.
  • Song Y; College of Veterinary Medicine, Henan Agricultural University, Zhengzhou 450046, China.
  • Ren H; Molecule Biology Laboratory, Zhengzhou Normal University, Zhengzhou 450044, China.
  • Zeng Q; College of Veterinary Medicine, Henan Agricultural University, Zhengzhou 450046, China.
  • Yuan Y; College of Veterinary Medicine, Henan Agricultural University, Zhengzhou 450046, China.
  • Xia L; College of Veterinary Medicine, Henan Agricultural University, Zhengzhou 450046, China.
  • Wei Z; College of Veterinary Medicine, Henan Agricultural University, Zhengzhou 450046, China.
Viruses ; 14(4)2022 04 08.
Article in English | MEDLINE | ID: covidwho-1810315
ABSTRACT
Porcine deltacoronavirus (PDCoV) mainly causes severe diarrhea and intestinal pathological damage in piglets and poses a serious threat to pig farms. Currently, no effective reagents or vaccines are available to control PDCoV infection. Single-chain fragment variable (scFv) antibodies can effectively inhibit virus infection and may be a potential therapeutic reagent for PDCoV treatment. In this study, a porcine phage display antibody library from the peripheral blood lymphocytes of piglets infected with PDCoV was constructed and used to select PDCoV-specific scFv. The library was screened with four rounds of biopanning using the PDCoV N protein, and the colony with the highest affinity to the PDCoV N protein was obtained (namely, N53). Then, the N53-scFv gene fragment was cloned into plasmid pFUSE-hIgG-Fc2 and expressed in HEK-293T cells. The scFv-Fc antibody N53 (namely, scFv N53) was purified using Protein A-sepharose. The reactive activity of the purified antibody with the PDCoV N protein was confirmed by indirect enzyme-linked immunosorbent assay (ELISA), western blot and indirect immunofluorescence assay (IFA). Finally, the antigenic epitopes that the scFv N53 recognized were identified by a series of truncated PDCoV N proteins. The amino acid residues 82GELPPNDTPATTRVT96 of the PDCoV N protein were verified as the minimal epitope that can be recognized by the scFv-Fc antibody N53. In addition, the interaction between the scFv-Fc antibody N53 and the PDCoV N protein was further analyzed by molecule docking. In conclusion, our research provides some references for the treatment and prevention of PDCoV.
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Full text: Available Collection: International databases Database: MEDLINE Main subject: Swine Diseases / Bacteriophages / Coronavirus Infections / Single-Chain Antibodies Topics: Vaccines Limits: Animals Language: English Year: 2022 Document Type: Article Affiliation country: V14040772

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Full text: Available Collection: International databases Database: MEDLINE Main subject: Swine Diseases / Bacteriophages / Coronavirus Infections / Single-Chain Antibodies Topics: Vaccines Limits: Animals Language: English Year: 2022 Document Type: Article Affiliation country: V14040772