PMID- 23314370
OWN - NLM
STAT- MEDLINE
DCOM- 20130619
LR  - 20190923
IS  - 1738-8872 (Electronic)
IS  - 1017-7825 (Linking)
VI  - 23
IP  - 1
DP  - 2013 Jan
TI  - Production and characterization of a monoclonal antibody against enrofloxacin.
PG  - 69-75
AB  - Enrofloxacin is a fluoroquinolone antibiotic approved for the treatment of 
      infections in animals. Because of the side effects to consumers of animal 
      products, the maximum residue limits (MRLs) of enrofloxacin in animal tissues for 
      consumption are regulated. In this study, a monoclonal antibody (mAb) against 
      enrofloxacin was prepared and characterized for the development of a direct 
      competitive enzyme-linked immunosorbent assay (ELISA). The obtained mAb, Enro44, 
      was highly specific for enrofloxacin and had a 50% inhibition concentration 
      (IC(50)) of 1.99 ng/ml in a competitive ELISA, and the limit of detection (LOD) 
      was 0.50 ng/ml. The cross-reactivity of the mAb with other quinolones and 
      fluoroquinolones was lower than 0.01%. The subclass of the mAb Enro44 was 
      identified as IgG1. The antigen (Ag)-captured direct competitive ELISA using the 
      mAb Enro44 was tested on different spiked samples, including chicken muscle, 
      cattle milk, and cattle urine, and the assay demonstrated recoveries of 82-112%, 
      80-125%, and 78-124%, respectively. Furthermore, the quantitation of enrofloxacin 
      obtained from the ELISA and from high-performance liquid chromatography (HPLC) 
      was in good agreement, with the linear regression coefficient between 0.933 and 
      1.056. The cDNAs encoding a heavy-chain Fd fragment (VH and CH1) and a light 
      chain of the mAb Enro44 were cloned and sequenced. Taken together, the results 
      obtained reveal a potential use of this mAb in an ELISA for the detection of 
      enrofloxacin in food samples. The information of amino acid sequence of this mAb 
      will be useful for further modification and production of the mAb in a 
      bioreactor.
FAU - Chusri, Manaspong
AU  - Chusri M
AD  - Program in Biotechnology, Faculty of Science, Chulalongkorn University, Bangkok 
      10330, Thailand.
FAU - Wongphanit, Pitikarn
AU  - Wongphanit P
FAU - Palaga, Tanapat
AU  - Palaga T
FAU - Puthong, Songchan
AU  - Puthong S
FAU - Sooksai, Sarintip
AU  - Sooksai S
FAU - Komolpis, Kittinan
AU  - Komolpis K
LA  - eng
SI  - GENBANK/JQ048936
SI  - GENBANK/JQ048937
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - Korea (South)
TA  - J Microbiol Biotechnol
JT  - Journal of microbiology and biotechnology
JID - 9431852
RN  - 0 (Antibodies, Monoclonal)
RN  - 0 (DNA, Complementary)
RN  - 0 (Fluoroquinolones)
RN  - 0 (Immunoglobulin G)
RN  - 3DX3XEK1BN (Enrofloxacin)
SB  - IM
MH  - Animals
MH  - Antibodies, Monoclonal/genetics/*immunology/isolation & purification
MH  - Cattle
MH  - *Chemistry Techniques, Analytical
MH  - Chickens
MH  - Cloning, Molecular
MH  - Cross Reactions
MH  - DNA, Complementary/genetics
MH  - Enrofloxacin
MH  - Enzyme-Linked Immunosorbent Assay/methods
MH  - Fluoroquinolones/*analysis/*immunology
MH  - Food Analysis/*methods
MH  - Immunoglobulin G/genetics/immunology/isolation & purification
MH  - Inhibitory Concentration 50
MH  - Molecular Sequence Data
MH  - Sensitivity and Specificity
MH  - Sequence Analysis, DNA
EDAT- 2013/01/15 06:00
MHDA- 2013/06/20 06:00
CRDT- 2013/01/15 06:00
PHST- 2013/01/15 06:00 [entrez]
PHST- 2013/01/15 06:00 [pubmed]
PHST- 2013/06/20 06:00 [medline]
AID - JMB023-01-11 [pii]
AID - 10.4014/jmb.1201.01017 [doi]
PST - ppublish
SO  - J Microbiol Biotechnol. 2013 Jan;23(1):69-75. doi: 10.4014/jmb.1201.01017.