PMID- 31420069
OWN - NLM
STAT- MEDLINE
DCOM- 20191104
LR  - 20191210
IS  - 1873-2534 (Electronic)
IS  - 0165-2427 (Linking)
VI  - 215
DP  - 2019 Sep
TI  - Generation of porcine monoclonal antibodies based on single cell technologies.
PG  - 109913
LID - S0165-2427(19)30109-6 [pii]
LID - 10.1016/j.vetimm.2019.109913 [doi]
AB  - The development of a rapid and efficient system to generate porcine monoclonal 
      antibodies (mAbs) is an important step toward the discovery of critical 
      neutralizing targets for designing rational vaccines against porcine viruses. In 
      this study, we established a platform for producing porcine mAbs based on single 
      cell technologies. First, we singled out an optimal donor from 507 pigs based on 
      serum antibody neutralizing activity against porcine reproductive and respiratory 
      syndrome virus (PRRSV). After identifying the contribution of IgG to the 
      neutralizing activity, single CD45R(+)IgG(+)Ag(+) B cells were sorted from 
      peripheral blood mononuclear cells (PBMCs). Single B cell RT-PCR was performed 
      using primers designed to cover the germline repertoire of the porcine VH/VL gene 
      segments. Paired VH/VLs were cloned into a eukaryotic expression vector and 
      transfected into 293T cells. We demonstrate that full-length porcine mAbs were 
      produced, and antigen-specific mAbs were obtained after further validation. The 
      approach reported in this study can be applied to generate porcine mAbs against 
      any given antigen and may help with the screening of neutralizing antibodies 
      against porcine pathogens.
CI  - Copyright (c) 2019 Elsevier B.V. All rights reserved.
FAU - Li, Kongpan
AU  - Li K
AD  - State Key Laboratory of Agrobiotechnology, College of Biological Sciences, 
      National Engineering Laboratory for Animal Breeding, China Agricultural 
      University, Beijing, People's Republic of China.
FAU - Bai, Jianhui
AU  - Bai J
AD  - State Key Laboratory of Agrobiotechnology, College of Biological Sciences, 
      National Engineering Laboratory for Animal Breeding, China Agricultural 
      University, Beijing, People's Republic of China.
FAU - Du, Lijuan
AU  - Du L
AD  - State Key Laboratory of Agrobiotechnology, College of Biological Sciences, 
      National Engineering Laboratory for Animal Breeding, China Agricultural 
      University, Beijing, People's Republic of China.
FAU - Wang, Xifeng
AU  - Wang X
AD  - Key Laboratory of Animal Ecology and Conservation Biology, Institute of Zoology, 
      Chinese Academy of Science, Beijing, People's Republic of China.
FAU - Ke, Cuncun
AU  - Ke C
AD  - State Key Laboratory of Agrobiotechnology, College of Biological Sciences, 
      National Engineering Laboratory for Animal Breeding, China Agricultural 
      University, Beijing, People's Republic of China.
FAU - Yan, Wei
AU  - Yan W
AD  - XINDAMUYE Company, Henan, People's Republic of China.
FAU - Li, Changqing
AU  - Li C
AD  - XINDAMUYE Company, Henan, People's Republic of China.
FAU - Ren, Liming
AU  - Ren L
AD  - State Key Laboratory of Agrobiotechnology, College of Biological Sciences, 
      National Engineering Laboratory for Animal Breeding, China Agricultural 
      University, Beijing, People's Republic of China.
FAU - Han, Haitang
AU  - Han H
AD  - State Key Laboratory of Agrobiotechnology, College of Biological Sciences, 
      National Engineering Laboratory for Animal Breeding, China Agricultural 
      University, Beijing, People's Republic of China. Electronic address: 
      hanhaitang@cau.edu.cn.
FAU - Zhao, Yaofeng
AU  - Zhao Y
AD  - State Key Laboratory of Agrobiotechnology, College of Biological Sciences, 
      National Engineering Laboratory for Animal Breeding, China Agricultural 
      University, Beijing, People's Republic of China. Electronic address: 
      yaofengzhao@cau.edu.cn.
LA  - eng
PT  - Journal Article
PT  - Validation Study
DEP - 20190731
PL  - Netherlands
TA  - Vet Immunol Immunopathol
JT  - Veterinary immunology and immunopathology
JID - 8002006
RN  - 0 (Antibodies, Monoclonal)
RN  - 0 (Antibodies, Neutralizing)
RN  - 0 (Antibodies, Viral)
RN  - 0 (Immunoglobulin Variable Region)
SB  - IM
MH  - Animals
MH  - Antibodies, Monoclonal/*biosynthesis/genetics/immunology/isolation & purification
MH  - Antibodies, Neutralizing/genetics/immunology
MH  - Antibodies, Viral/*biosynthesis/genetics/immunology/isolation & purification
MH  - Antibody Specificity
MH  - B-Lymphocytes/immunology
MH  - HEK293 Cells
MH  - Humans
MH  - Immunoglobulin Variable Region/genetics
MH  - Porcine respiratory and reproductive syndrome virus/*immunology
MH  - Real-Time Polymerase Chain Reaction/veterinary
MH  - Swine/*immunology
MH  - Transfection
MH  - V(D)J Recombination
OTO - NOTNLM
OT  - Antigen-specific B cell
OT  - PRRSV
OT  - Porcine monoclonal antibody
OT  - Single cell RT-PCR
EDAT- 2019/08/20 06:00
MHDA- 2019/11/05 06:00
CRDT- 2019/08/18 06:00
PHST- 2019/03/28 00:00 [received]
PHST- 2019/07/17 00:00 [revised]
PHST- 2019/07/30 00:00 [accepted]
PHST- 2019/08/18 06:00 [entrez]
PHST- 2019/08/20 06:00 [pubmed]
PHST- 2019/11/05 06:00 [medline]
AID - S0165-2427(19)30109-6 [pii]
AID - 10.1016/j.vetimm.2019.109913 [doi]
PST - ppublish
SO  - Vet Immunol Immunopathol. 2019 Sep;215:109913. doi: 10.1016/j.vetimm.2019.109913. 
      Epub 2019 Jul 31.