PMID- 38143067
OWN - NLM
STAT- MEDLINE
DCOM- 20240208
LR  - 20240208
IS  - 1879-0003 (Electronic)
IS  - 0141-8130 (Linking)
VI  - 258
IP  - Pt 2
DP  - 2024 Feb
TI  - Multivalent nanobody-based sandwich enzyme-linked immunosorbent assay for 
      sensitive detection of porcine reproductive and respiratory syndrome virus.
PG  - 128896
LID - S0141-8130(23)05795-1 [pii]
LID - 10.1016/j.ijbiomac.2023.128896 [doi]
AB  - The pandemic of the porcine reproductive and respiratory syndrome virus (PRRSV) 
      has caused huge economic losses and continues to threaten the swine industry 
      worldwide. Nucleocapsid protein (N protein) is the primary antigen of PRRSV for 
      development of sensitive diagnostic assays. Two high affinity nanobodies against 
      N protein, Nb12 and Nb35, were selected and employed to develop a sandwich ELISA. 
      Further we improved the ELISA method to obtain greater sensitivity, a trivalent 
      nanobody (3 x Nb35) and a bivalent nanobody-HRP fusion protein (2 x Nb12-HRP) 
      were expressed and used. This modified ELISA was found to have high sensitivity 
      for detecting PRRSV, with a detection limit of 10 TCID(50)/ml (median tissue 
      culture infectious dose), which was approximately 200-fold greater than the 
      single-copy nanobody-based sandwich ELISA. The developed assay shows high 
      specificity and can detect almost all circulating lineages of PRRSV-2 in China. 
      This study provides suggestions for reforming nanobodies and for the further 
      development of multivalent nanobody-based ELISAs for other various viruses.
CI  - Copyright (c) 2023. Published by Elsevier B.V.
FAU - Sun, Mingxia
AU  - Sun M
AD  - State Key Laboratory for Animal Disease Control and Prevention, Harbin Veterinary 
      Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150069, 
      China.
FAU - Sun, Yue
AU  - Sun Y
AD  - State Key Laboratory for Animal Disease Control and Prevention, Harbin Veterinary 
      Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150069, 
      China.
FAU - Yang, Yongbo
AU  - Yang Y
AD  - State Key Laboratory for Animal Disease Control and Prevention, Harbin Veterinary 
      Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150069, 
      China.
FAU - Zhao, Man
AU  - Zhao M
AD  - State Key Laboratory for Animal Disease Control and Prevention, Harbin Veterinary 
      Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150069, 
      China.
FAU - Cao, Dan
AU  - Cao D
AD  - Soybean Research Institute, Heilongjiang Academy of Agricultural Science, Harbin 
      150086, China.
FAU - Zhang, Minmin
AU  - Zhang M
AD  - State Key Laboratory for Animal Disease Control and Prevention, Harbin Veterinary 
      Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150069, 
      China.
FAU - Xia, Dasong
AU  - Xia D
AD  - State Key Laboratory for Animal Disease Control and Prevention, Harbin Veterinary 
      Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150069, 
      China.
FAU - Wang, Tao
AU  - Wang T
AD  - State Key Laboratory for Animal Disease Control and Prevention, Harbin Veterinary 
      Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150069, 
      China.
FAU - Gao, Yanfei
AU  - Gao Y
AD  - State Key Laboratory for Animal Disease Control and Prevention, Harbin Veterinary 
      Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150069, 
      China.
FAU - Wang, Shanghui
AU  - Wang S
AD  - State Key Laboratory for Animal Disease Control and Prevention, Harbin Veterinary 
      Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150069, 
      China.
FAU - Wang, Haiwei
AU  - Wang H
AD  - State Key Laboratory for Animal Disease Control and Prevention, Harbin Veterinary 
      Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150069, 
      China.
FAU - Cai, Xuehui
AU  - Cai X
AD  - State Key Laboratory for Animal Disease Control and Prevention, Harbin Veterinary 
      Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150069, 
      China; Heilongjiang Research Center of Veterinary Biopharmaceutical Technology, 
      Harbin 150069, China. Electronic address: caixuehui@caas.cn.
FAU - An, Tongqing
AU  - An T
AD  - State Key Laboratory for Animal Disease Control and Prevention, Harbin Veterinary 
      Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150069, 
      China; Heilongjiang Provincial Key Laboratory of Veterinary Immunology, Harbin 
      150069, China. Electronic address: antongqing@caas.cn.
LA  - eng
PT  - Journal Article
DEP - 20231222
PL  - Netherlands
TA  - Int J Biol Macromol
JT  - International journal of biological macromolecules
JID - 7909578
RN  - 0 (Single-Domain Antibodies)
RN  - 0 (Antigens, Viral)
RN  - 0 (Nucleocapsid Proteins)
RN  - 0 (Antibodies, Viral)
SB  - IM
MH  - Animals
MH  - Swine
MH  - *Porcine respiratory and reproductive syndrome virus
MH  - *Porcine Reproductive and Respiratory Syndrome
MH  - *Single-Domain Antibodies/metabolism
MH  - Enzyme-Linked Immunosorbent Assay/methods
MH  - Antigens, Viral
MH  - Nucleocapsid Proteins
MH  - Antibodies, Viral
MH  - Sensitivity and Specificity
OTO - NOTNLM
OT  - Multivalent nanobody
OT  - Nanobody-HRP
OT  - PRRSV
OT  - Sandwich ELISA
COIS- Declaration of competing interest The authors declare that they have no competing 
      interests.
EDAT- 2023/12/25 00:42
MHDA- 2024/02/08 06:43
CRDT- 2023/12/24 19:32
PHST- 2023/08/01 00:00 [received]
PHST- 2023/10/13 00:00 [revised]
PHST- 2023/12/17 00:00 [accepted]
PHST- 2024/02/08 06:43 [medline]
PHST- 2023/12/25 00:42 [pubmed]
PHST- 2023/12/24 19:32 [entrez]
AID - S0141-8130(23)05795-1 [pii]
AID - 10.1016/j.ijbiomac.2023.128896 [doi]
PST - ppublish
SO  - Int J Biol Macromol. 2024 Feb;258(Pt 2):128896. doi: 
      10.1016/j.ijbiomac.2023.128896. Epub 2023 Dec 22.