Synthesis and characterization of hapten-protein conjugates for antibody production against small molecules.

Singh, K V and Kaur, Jasdeep and Varshney, Grish C and Raje, Manoj and Suri, C Raman (2004) Synthesis and characterization of hapten-protein conjugates for antibody production against small molecules. Bioconjugate chemistry, 15 (1). pp. 168-73. ISSN 1043-1802

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Abstract

For the generation of antibodies against small hapten molecules, the hapten is cross-linked with some carrier protein to make it immunogenic. However, the formation of such conjugates is not always reproducible. This may lead to inconsistent hapten-protein stoichiometries, resulting in large variations in the generation of the desired antibodies. In the study described here the hapten (mercaptopropionic acid derivative of atrazine) was coupled to carrier protein at five different molar ratios. The hapten-protein conjugates prepared were characterized thoroughly by spectrophotometric absorption, fluorescence, matrix-assisted laser desorption ionization (MALDI), and gel electrophoresis methods, before being used for the immunization and assay purposes. Electrophoresis and fluorescence methods were very useful in detecting hapten-protein cross-linking while MALDI-MS and spectrophotometric detection provided qualitatively comparable hapten density. The production of specific antibodies was sought following the generation of appropriate hapten-protein conjugates. A high antibody titer with moderate antibody specificity was obtained with hapten density around 15 molecules per carrier protein. The study proved useful for monitoring the course of hapten-protein conjugation for the production of specific antibodies against small molecules.

Item Type:	Article
Additional Information:	Copyright of this article belongs to ACS.
Subjects:	Q Science > QD Chemistry
Depositing User:	Dr. K.P.S.Sengar
Date Deposited:	06 Jan 2012 14:55
Last Modified:	07 Jan 2012 10:51
URI:	http://crdd.osdd.net/open/id/eprint/224

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