PMID- 22533788
OWN - NLM
STAT- MEDLINE
DCOM- 20120914
LR  - 20120523
IS  - 1520-5118 (Electronic)
IS  - 0021-8561 (Linking)
VI  - 60
IP  - 20
DP  - 2012 May 23
TI  - Production and characterization of a single-chain variable fragment linked 
      alkaline phosphatase fusion protein for detection of O,O-diethyl organophosphorus 
      pesticides in a one-step enzyme-linked immunosorbent assay.
PG  - 5076-83
LID - 10.1021/jf300570q [doi]
AB  - A single-chain variable fragment (scFv) linked alkaline phosphatase (AP) fusion 
      protein for detection of O,O-diethyl organophosphorus pesticides (O,O-diethyl 
      OPs) was produced and characterized. The scFv gene was prepared by cloning V(L) 
      and V(H) genes from hybridoma cells secreting monoclonal antibody with broad 
      specificity for O,O-diethyl OPs. The amplified V(L) and V(H) regions were 
      assembled using a linker (Gly(4)Ser)(3) by means of splicing overlap extension 
      polymerase chain reaction to obtain the scFv gene, which was cloned into the 
      expression vector pLIP6/GN containing an AP gene to produce the scFv-AP fusion 
      protein in Escherichia coli strain BL21. The protein was purified by 
      antigen-conjugated immunoaffinity chromatography and characterized by sodium 
      dodecyl sulfate-polyacrylamide gel electrophoresis, Western blotting, and 
      competitive direct enzyme-linked immunosorbent assay (cdELISA). The fusion 
      protein is bifunctional, retaining both antigen binding specificity and AP 
      enzymatic activity. Analysis of spiked and blind river water and Chinese cabbage 
      samples demonstrated that the fusion protein based cdELISA(FP) exhibited good 
      sensitivity and reproducibility.
FAU - Xu, Zhen-Lin
AU  - Xu ZL
AD  - Guangdong Provincial Key Laboratory of Food Quality and Safety, South China 
      Agricultural University , Guangzhou 510642, China.
FAU - Dong, Jie-Xian
AU  - Dong JX
FAU - Wang, Hong
AU  - Wang H
FAU - Li, Zhen-Feng
AU  - Li ZF
FAU - Beier, Ross C
AU  - Beier RC
FAU - Jiang, Yue-Ming
AU  - Jiang YM
FAU - Lei, Hong-Tao
AU  - Lei HT
FAU - Shen, Yu-Dong
AU  - Shen YD
FAU - Yang, Jin-Yi
AU  - Yang JY
FAU - Sun, Yuan-Ming
AU  - Sun YM
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20120514
PL  - United States
TA  - J Agric Food Chem
JT  - Journal of agricultural and food chemistry
JID - 0374755
RN  - 0 (Antibodies, Monoclonal)
RN  - 0 (Organophosphorus Compounds)
RN  - 0 (Pesticides)
RN  - 0 (Recombinant Fusion Proteins)
RN  - 0 (Single-Chain Antibodies)
RN  - EC 3.1.3.1 (Alkaline Phosphatase)
SB  - IM
MH  - *Alkaline Phosphatase
MH  - Amino Acid Sequence
MH  - Antibodies, Monoclonal/immunology
MH  - Antibody Specificity
MH  - Base Sequence
MH  - Cloning, Molecular
MH  - Enzyme-Linked Immunosorbent Assay/*methods
MH  - Food Contamination/analysis
MH  - Hybridomas/immunology
MH  - Molecular Sequence Data
MH  - Organophosphorus Compounds/*analysis
MH  - Pesticides/*analysis
MH  - *Recombinant Fusion Proteins/genetics/immunology
MH  - Reproducibility of Results
MH  - Sensitivity and Specificity
MH  - *Single-Chain Antibodies/chemistry/genetics/immunology
EDAT- 2012/04/27 06:00
MHDA- 2012/09/15 06:00
CRDT- 2012/04/27 06:00
PHST- 2012/04/27 06:00 [entrez]
PHST- 2012/04/27 06:00 [pubmed]
PHST- 2012/09/15 06:00 [medline]
AID - 10.1021/jf300570q [doi]
PST - ppublish
SO  - J Agric Food Chem. 2012 May 23;60(20):5076-83. doi: 10.1021/jf300570q. Epub 2012 
      May 14.