Escherichia coli expressing single-chain Fv on the cell surface as a potential prophylactic of porcine epidemic diarrhea virus.
Porcine epidemic diarrhea virus (PEDV) is a causative agent of severe diarrhea which leads to death in piglets. Because of the high mortality which is up to 100% in suckling piglets, PED is an important porcine disease in Korea. In this study, we developed a prophylactic candidate using single-chain Fvs to prevent the PEDV infection. ScFvs of mouse monoclonal antibody which was verified to neutralize PEDV was expressed in Escherichia coli expression system. After the confirmation of PEDV neutralizing activity of purified recombinant scFvs by VN test, scFvs were expressed on the surface of E. coli cells. The signal sequence and autotransporter beta domain of protease IgA (IgAP) of Neisseria gonorrhoeae were introduced to endow scFvs with the direction to the cell surface and the support as a transmembrane domain. 5x10(6)CFU of E. coli expressing scFvs against PEDV showed promising result of 94% foci reduction compared to wild type E. coli. This result demonstrated that E. coli expressing scFvs on the cell surface retained functional potency of parent antibody and therefore blocked PEDV infection into target cells in vitro. This in vitro assay result proposes the perspective of recombinant E. coli cells expressing scFvs as a novel prophylactic against PEDV infection.
Pyo
Hyun-Mi
HM
National Veterinary Research and Quarantine Service, Ministry for Food, Agriculture, Forestry and Fisheries, Anyang 430-824, Republic of Korea.
Kim
In-Joong
IJ
Kim
Seong-Hee
SH
Kim
Hyun-Soo
HS
Cho
Soo-Dong
SD
Cho
In-Soo
IS
Hyun
Bang-Hun
BH
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D000918:Antibody Specificity; D018352:Coronavirus Infections / Q000517:prevention & control*; D003429:Cross Reactions; D003841:Deoxycytidine / Q000235:genetics; D000939:Epitopes / Q000276:immunology; D004926:Escherichia coli / Q000235:genetics / Q000276:immunology*; D007128:Immunoglobulin Fragments / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D008969:Molecular Sequence Data; D009500:Neutralization Tests; D053485:Porcine epidemic diarrhea virus / Q000276:immunology*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D012697:Serine Endopeptidases / Q000276:immunology; D013553:Swine Diseases / Q000517:prevention & control* / Q000821:virology
D000911:Antibodies, Monoclonal; D000939:Epitopes; D007128:Immunoglobulin Fragments; D011994:Recombinant Proteins; D003841:Deoxycytidine; D012697:Serine Endopeptidases; C021433:IgA-specific serine endopeptidase