Screening scFv antibodies against infectious bursal disease virus by co-expression of antigen and antibody in the bacteria display system.
We have previously reported an antigen and antibody co-expression (AAC) technology to demonstrate the interaction between a known antigen and antibody. To validate the co-expression system for screening antibody libraries, a single chain fragment variable(scFv)antibody library was constructed from chickens immunized with the VP2 protein of infectious bursal disease virus (IBDV). The genes of both VP2 and scFv antibodies were inserted into the pBFD-Ab-Ag vector. The co-expression library was subjected to three rounds of screening by flow cytometry (FCM) using a polyclonal antibody against VP2 through a bacteria display system. We achieved enrichment of scFv specific for IBDV. 110 individual clones were initially selected and sequenced. Twenty clones were selected based on fluorescence intensity by FCM. The scFv antibodies were expressed by pET-27b in E.coli and purified. The specificity and affinity of the selected scFv antibodies were confirmed by western blotting assay and ELISA analysis. What's more, the neutralizing capacity was measured with IBDV-B87(100 TCID50) in vitro. Four scFvs (clone 8(1), Y8, L10 and L7) showed significant neutralizing capacity. Two of the four scFvs (clone 8(1) and Y8) demonstrated a higher neutralizing activity to IBDV-B87 and the titers were 16,384 and 8,192, respectively. The two scFvs had higher neutralizing capacity than those obtained in previous studies. We demonstrated that the AAC technology could be applied to screen antibody libraries without baiting antigen to make antibody screening process easier and obtain scFvs with higher neutralizing capacity.
Biopharmaceutical Lab, College of Life Science, Northeast Agricultural University, Harbin 150030, China.
Xiaochen
X
Guo
Biopharmaceutical Lab, College of Life Science, Northeast Agricultural University, Harbin 150030, China.
Hongxue
H
Cao
Biopharmaceutical Lab, College of Life Science, Northeast Agricultural University, Harbin 150030, China.
Yunxin
Y
Wang
Biopharmaceutical Lab, College of Life Science, Northeast Agricultural University, Harbin 150030, China.
Yang
Y
Liu
Biopharmaceutical Lab, College of Life Science, Northeast Agricultural University, Harbin 150030, China.
Yanmin
Y
Chen
Biopharmaceutical Lab, College of Life Science, Northeast Agricultural University, Harbin 150030, China.
Nan
N
Wang
Biopharmaceutical Lab, College of Life Science, Northeast Agricultural University, Harbin 150030, China.
Shan
S
Jiang
Biopharmaceutical Lab, College of Life Science, Northeast Agricultural University, Harbin 150030, China.
Shengqi
S
Zhang
Biopharmaceutical Lab, College of Life Science, Northeast Agricultural University, Harbin 150030, China.
Qiang
Q
Wu
Beijing Obstetrics and Gynecology Hospital, Capital Medical University, Beijing 100006, China.
Tianhe
T
Li
Biopharmaceutical Lab, College of Life Science, Northeast Agricultural University, Harbin 150030, China.
Yingjie
Y
Zhang
National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Public Health, Xiamen University, Xiamen 361102, China.
Bing
B
Zhou
Biopharmaceutical Lab, College of Life Science, Northeast Agricultural University, Harbin 150030, China.
Jiechao
J
Yin
Biopharmaceutical Lab, College of Life Science, Northeast Agricultural University, Harbin 150030, China; Key Laboratory of Agricultural Biological Functional Gene, Northeast Agricultural University, Harbin 150030, China.
Deshan
D
Li
Biopharmaceutical Lab, College of Life Science, Northeast Agricultural University, Harbin 150030, China; Key Laboratory of Agricultural Biological Functional Gene, Northeast Agricultural University, Harbin 150030, China. Electronic address: renguiping@126.com.
Guiping
G
Ren
D000818:Animals; D000937:Antigen-Antibody Reactions; D001419:Bacteria / Q000235:genetics; D002645:Chickens; D005434:Flow Cytometry; D007128:Immunoglobulin Fragments / Q000235:genetics* / Q000276:immunology; D007243:Infectious bursal disease virus / Q000276:immunology*; D009500:Neutralization Tests; D019151:Peptide Library; D057127:Single-Chain Antibodies / Q000235:genetics* / Q000276:immunology
D007128:Immunoglobulin Fragments; D019151:Peptide Library; D057127:Single-Chain Antibodies; C055966:immunoglobulin Fv
Antibody library; Antigen and antibody co-expression technology; Bacterial display; IBDV; scFv.