Selection and characterization of scFv antibody against nucleocapsid protein of Porcine reproductive and respiratory syndrome virus

Krásna, Magdaléna; Celer, Vladimír

doi:10.2754/avb202089010039

Acta Veterinaria > Archive > 2020, Vol. 89 > Issue 1 > Selection and characterization of scFv…

Acta Vet. Brno 2020, 89: 39-45

https://doi.org/10.2754/avb202089010039

Selection and characterization of scFv antibody against nucleocapsid protein of Porcine reproductive and respiratory syndrome virus

Magdaléna Krásna^1,2, Vladimír Celer^1,2

¹CEITEC – Central European Institute of Technology, Brno, Czech Republic
²University of Veterinary and Pharmaceutical Sciences Brno, Faculty of Veterinary Medicine, Institute of Infectious Diseases and Microbiology, Brno, Czech Republic

Received January 21, 2019
Accepted February 25, 2020

Porcine reproductive and respiratory syndrome virus (PRRSV) is a widespread infectious agent in pigs. Nucleocapsid (N) protein of PRRSV has been identified as the most immunodominant viral protein. The main goal of the work was the selection and characterization of a single-chain antibody fragments (scFv) antibody specific to the N protein. Specific scFv antibody clone D5 was selected from the Tomlinson phagemid library and purified by immobilized metal affinity chromatography from the periplasmatic space of E. coli cells. The antibody was then characterized by sequencing and the ability to recognize the native virus N protein by Western blot and competitive ELISA. Pepscan analysis identified the position of the binding epitope between amino acids 62–84 of the N protein. Our study could help to improve the diagnostics and prevention of PRRSV in Central Europe.

Keywords

N protein, expression, binding epitope, immunoblot, ELISA.

Funding

This work was supported by “CEITEC – Central European Institute of Technology” (CZ.1.05/1.1.00/02.0068) from the European Regional Development Fund and the project QJ1510108 funded by National Agency for Agricultural Research (NAZV), Czech Republic.