Monoclonal antibody to N protein of porcine epidemic diarrhea virus.
The N gene of porcine epidemic diarrhea virus (PEDV) was amplified by RT-PCR using specific primers, and inserted into the expression vector pCold-I to construct a recombinant plasmid pCold-I-N. The recombinant plasmid was expressed in Escherichia coli BL21 (DE3) under IPTG induction. Then, female BALB/c mice were immunized with the purified recombinant N protein and one strain of hybridoma cells named 2B8 secreting anti-N protein monoclonal antibodies (MAb) was obtained by hybridoma technique. The MAb was specifically reacted with PEDV and identified by Western blot and indirect immunofluorescence assays. This work indicated that the MAb would be a valuable tool as a specific diagnostic reagent for PEDV epidemiological surveys and diagnosis in the future.
1 Department of Swine Infectious Disease, Shanghai Veterinary Research Institute , Chinese Academy of Agricultural Sciences, Shanghai, China .
Xi
X
Pan
Ning
N
Kong
Tongling
T
Shan
Hao
H
Zheng
Wu
W
Tong
Shen
S
Yang
Guoxin
G
Li
Enmin
E
Zhou
Guangzhi
G
Tong
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000302:isolation & purification; D000914:Antibodies, Viral / Q000096:biosynthesis* / Q000302:isolation & purification; D000918:Antibody Specificity; D015153:Blotting, Western; D002460:Cell Line; D018352:Coronavirus Infections / Q000175:diagnosis* / Q000276:immunology / Q000662:veterinary* / Q000821:virology; D000086462:Coronavirus Nucleocapsid Proteins; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005260:Female; D019084:Fluorescent Antibody Technique, Indirect; D018703:Founder Effect; D015870:Gene Expression; D006825:Hybridomas / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D019590:Nucleocapsid Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D053485:Porcine epidemic diarrhea virus / Q000276:immunology / Q000302:isolation & purification*; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D013552:Swine; D013553:Swine Diseases / Q000175:diagnosis* / Q000276:immunology / Q000821:virology
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000086462:Coronavirus Nucleocapsid Proteins; D019590:Nucleocapsid Proteins; D011994:Recombinant Proteins