Title :
Preliminary Production and Characterization of Anti-Monocrotophos Monoclonal Antibody
Author :
Xia Ying ; Yu Xiao-Wei ; Yang Di ; Yuan Jun-lin
Author_Institution :
Coll. of Life Sci., Huazhong Normal Univ., Wuhan, China
Abstract :
Monocrotophos as the hapten was coupled the carrier proteins ovalbumin (OVA) and bovine serum albumin (BSA) by Glutaral Method to synthesis artificial antigen and coating antigen. The identifications of synthetic antigens were by ultraviolet absorption spectrum scan and SDS-PAGE methods. And then BALB/C mices were immunized with immunogen (hapten-OVA). The result showed that UV spectrogram of synthetic antigens was changed when compared with OVA and BSA, electrophoretic mobility between synthetic antigens and carrier proteins was different. The coupling molar ratios of the hapten and carrier proteins were 23.3:1 for hapten-OVA, 13.7:1 for hapten-BSA. Through examinations, the titer of the antiserum of mice is 1:409600. It could be concluded that Monocrotophos artificial antigen was synthesized and mice were immunized successfully. After cell fusion and cloned strain was established, the characterization of antibody was identified, the results indicated: the subclass was IgM, the molecular weight was 100.6KDa, and the chromosomal numbers were between 97 and 104. With large-scale preparation of this monoclonal antibody, it was purified and analyzed by SDS-PAGE. That provided a basis for further establishing immunological assay detection and for Monocrotophos residue and related performances on pesticide studies.
Keywords :
biological techniques; cellular biophysics; electrophoresis; molecular biophysics; proteins; ultraviolet spectra; anti-monocrotophos monoclonal antibody; carrier protein ovalbumin; carrier proteins; cell fusion; chromosomal numbers; cloned strain; electrophoretic mobility; immunological assay detection; mice antiserum; molecular weight; monocrotophos artificial antigen; pesticide; synthetic antigens; ultraviolet absorption spectrum; Biological cells; Bovine; Capacitive sensors; Coatings; Electromagnetic wave absorption; Large-scale systems; Mice; Production; Proteins; Spectrogram;
Conference_Titel :
Bioinformatics and Biomedical Engineering , 2009. ICBBE 2009. 3rd International Conference on
Conference_Location :
Beijing
Print_ISBN :
978-1-4244-2901-1
Electronic_ISBN :
978-1-4244-2902-8
DOI :
10.1109/ICBBE.2009.5163219