Screening of anti-human albumin monoclonal antibody by chemiluminescence immunoassay
Abstract
In this study, chemiluminescence immunoassay was used to screen clones of mouse anti-human albumin monoclonal antibody (anti-ALB McAb). The method is simplified as a one-step operation and the experimental parameters are optimized by orthogonal experiment [L9 (34)]. The parameters of the positive clone / blank control luminescence value (SNR) are the largest and the method is evaluated and statistical analysis. The results showed that the coated antibody was 3mg / L, the ratio of the enzyme protein A was 1: 2000, the incubation time was 304 times and the signal - to - noise ratio was 1284, which was several times higher than other combinations. The linear range was 20-20000ng / L, the precision was good, the average CV was 5.32% and the average inter-assay CV was 8.82%. The results showed that the method can be used for cloning and screening of anti-ALB McAb and the method is obviously superior to ELISA and other traditional methods, especially the simple and rapid operation of one step is also suitable for high antibody screening of other antibodies.References
Simons FE, Frew AJ, Ansotegui IJ, et al. Risk assessment in anaphylax-is: current and future approaches [J]. Allergy Clin
Immunol, 2007, 120 (1): 2-24.
G. C. Howard, M. R. Kaiser. Zhang Weigeng, Zhang Yuxiang, Ding Wei, et al., translations. Antibody preparation and use of
experimental guidelines [M]. Beijing, Science Press, 2010, 123-125.
Peavy RD, Metcalfe DD. Understanding the mechanisms of anaphylaxis [J]. Curr Opin Allergy Clin Immunol, 2008, 8 (4):
-315.
Zhang Zhengrong, Sun Hao, Lu Runming, et al. Study on enzyme-linked immunosorbent assay of recombinant antigens of
Treponema pallidum gene [J] Chinese Journal of Microbiology and Immunology, 2002,20 (1): 781.
Szebeni. Complement activation-related pseudoallergy: A new class of drug-induced acute fi re [J]. Toxicology, 2005, 216 (2-3):
-121.
Erik E, Marianne HH. Mast cell tryptase and hemolysis after trauma [J]. Forensic Science International, 2003, 131 (1): 8-13.
He S. Key role of mast cells and their major secretory products in infl ammatory bowel disease [J]. World J Gastroenterol,
,10 (3): 309-318.
Carvalho L, Largua A. The use of an automated chemiluminescent assay for screening of syphilis diagnostic [J]. Clin Chem
Lab Med, 2008, 46: 339-340.
Zhu Liping, Chen Xueqing, editor. Immunology commonly used experimental method [J]. Beijing, People's Medical Publishing
House, 2003.454.
Lin Jinming. Application of basic theory of chemiluminescence [M]. Beijing; Chemical Industry Press, 2004.125-127.
Wang will string. Production technology and application prospect of melamine [J]. Fine and Specialty Chemicals, 2007,15 (11):
-36.
Hu Guomao, Hu Zhangxue. Quantitative detection of hepatitis B virus markers by chemiluminescence immunoassay [J].
Labeled and Clinical Immunoassay, 2004, 113: 157-159.
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