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United States Department of Agriculture

Agricultural Research Service

Research Project: INTERVENTIONS TO REDUCE FOODBORNE PATHOGENS IN SWINE AND CATTLE

Location: Food and Feed Safety Research

Title: Monoclonal antibodies with group specificity toward sulfonamides: Selection of hapten and antibody selectivity

Authors
item Wang, Zhanhui -
item Beier, Ross
item Sheng, Yajie -
item Zhang, Suxia -
item Jiang, Wenxiao -
item Wang, Zhaopeng -
item Wang, Jin -
item Shen, Jianzhong -

Submitted to: Analytical and Bioanalytical Chemistry
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: January 24, 2013
Publication Date: February 16, 2013
Citation: Wang, Z., Beier, R.C., Sheng, Y., Zhang, S., Jiang, W., Wang, Z., Wang, J., Shen, J. 2013. Monoclonal antibodies with group specificity toward sulfonamides: Selection of hapten and antibody selectivity. Analytical and Bioanalytical Chemistry. 405:4027-4037.

Interpretive Summary: Antibodies (MAbs) are substances that are produced by the immune system in response to foreign substances that enter the body. When antibodies are isolated, they can be used as a diagnostic tool to detect foreign substances using a test called an ELISA (enzyme-linked immunosorbent assay). In this study, we developed new MAbs that are more efficient at detecting certain sulfanomide antibiotics than those currently being used. These new MAbs will improve our ability to detect certain sulfanomide antibiotics in animal tissue.

Technical Abstract: Although many antibodies to sulfonamides have been generated, immunoassays based on the current available antibodies for large multi-sulfonamide screening programs have properties dependent on the immunizing hapten structure and have always suffered from high selectivity for individual sulfonamides. In this study, two types of haptens (a total of seven) which may or may not contain a ring structure at the N1 position of the sulfonamides were selected to evaluate the effectiveness for producing group-specific monoclonal antibodies (MAbs) against sulfonamides. Mice immunized with three different two-ring haptens were used for hybridoma production, which resulted in three unique MAbs with excellent generic selectivity for sulfonamides, recognizing 10, 13, and 15 sulfonamides showing 50% inhibition (IC50) at concentrations below 100 ng/mL. Among them, the MAb 4D11 derived from one novel immunizing hapten could recognize 12 sulfonamides with IC50 values ranging from 1.2 to 12.4 ng/mL, almost within 1 order of magnitude. These produced MAbs, compared to previously generated MAbs, show better sensitivity based on much lower IC50 values in addition to significantly improved group selectivity with a narrow detectability interval. This study clearly indicates that the careful selection of the immunizing hapten has an important effect on the selectivity of the generated antibodies.

Last Modified: 7/25/2014
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