Recombinant scFv antibodies against infectious pancreatic necrosis virus isolated by flow cytometry.
Infectious pancreatic necrosis is a significant disease of farmed salmonids in China. In this study, a single chain variable fragment (scFv) antibody library derived from rainbow trout (Oncorhynchus mykiss) and viral protein VP2 of a Chinese infectious pancreatic necrosis virus (IPNV) isolate ChRtm213 were co-expressed by a bacterial display technology. The library was subjected to three rounds of screening by flow cytometry (FCM) to select IPNV specific antibodies. Six antibody clones with different mean fluorescence intensities (MFI) were obtained by picking colonies at random. The antibody clones were expressed and purified. The purified IPNV-specific scFv antibodies were used successfully in Western blotting, enzyme linked immunosorbent assay (ELISA) and an immunofluorescence antibody test (IFAT). This method provides a high throughput means to screen an antibody library by flow cytometry, and isolate a panel of antibody that can be used as potential reagents for the detection and study of IPNV that are prevalent in China.
Heilongjiang River Fishery Research Institute Chinese Academy of Fishery Sciences, Harbin 150070, PR China. Electronic address: 279463437@qq.com.
Li-Ming
LM
Xu
Heilongjiang River Fishery Research Institute Chinese Academy of Fishery Sciences, Harbin 150070, PR China. Electronic address: 422842454@qq.com.
Jing-Zhuang
JZ
Zhao
Heilongjiang River Fishery Research Institute Chinese Academy of Fishery Sciences, Harbin 150070, PR China. Electronic address: 446048847@qq.com.
Miao
M
Liu
Heilongjiang River Fishery Research Institute Chinese Academy of Fishery Sciences, Harbin 150070, PR China. Electronic address: 375825652@qq.com.
Yong-Sheng
YS
Cao
Heilongjiang River Fishery Research Institute Chinese Academy of Fishery Sciences, Harbin 150070, PR China. Electronic address: xwsc20@tom.com.
Jia-Sheng
JS
Yin
Heilongjiang River Fishery Research Institute Chinese Academy of Fishery Sciences, Harbin 150070, PR China. Electronic address: 112893129@qq.com.
Hong-Bai
HB
Liu
Heilongjiang River Fishery Research Institute Chinese Academy of Fishery Sciences, Harbin 150070, PR China. Electronic address: lutongyan@hotmail.com.
Tongyan
T
Lu
D000818:Animals; D000914:Antibodies, Viral / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D000915:Antibody Affinity; D018175:Birnaviridae Infections / Q000175:diagnosis / Q000276:immunology / Q000662:veterinary* / Q000821:virology; D002681:China / Q000453:epidemiology; D004797:Enzyme-Linked Immunosorbent Assay; D005393:Fish Diseases / Q000175:diagnosis / Q000276:immunology / Q000821:virology; D005434:Flow Cytometry / Q000379:methods*; D005453:Fluorescence; D015723:Gene Library; D057166:High-Throughput Screening Assays; D007158:Immunologic Techniques; D018061:Infectious pancreatic necrosis virus / Q000276:immunology*; D012474:Salmon / Q000821:virology; D057127:Single-Chain Antibodies / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*
D000914:Antibodies, Viral; D057127:Single-Chain Antibodies
Antibody library; Antibody-antigen co-expression; Bacterial display; Fluorescence activated cell sorter; Infectious pancreatic necrosis virus ChRtm213; Single chain variable fragment