SBIR-STTR Award

Neogen is committed to development of diagnostics which will improve the health of both plants and animals. Using information gathered from production of antibodies which recognize various mycotoxins, similar procedures would be used to produce polyclonal antiserum in rabbits and monoclonal antibodies (secreted by stabilized hybridoma cell lines) which bind to sulfonamides. The incorporation of antibiotics and the sulfonamides in livestock and poultry feeds has risen steadily since 1950, as animal production has expanded and grown. The concern over the addition of subtherapeutic doses of antimicrobials to animal feed is growing because of their potential as a human health hazard. Current methods for analysis of sulfonamides are labor-intensive, time-consuming, and can require extensive sample cleanup. They also detect sulfa residues at or near the tolerance level of 0.1 ppm. The goals of this research would be: (I) production of sensitive antibodies which could be used as analytical reagents to improve the assay time and limit of detectability of sulfonamides in samples requiring little or no cleanup prior to analysis, (2) successful production of stabilized cell lines which secrete sensitive antibodies would eliminate the potential variation observed from batch to batch of polyclonal antibody. A homogeneous, infinite supply of antibody could be obtained for use in kit production.Anticipated Results and

Potential Commercial Applications:
The antibodies would be used to develop an enzyme immunoassay for rapid detection of sulfonamides in clinically and agriculturally related samples. Similar technology has already been successfully implemented in detection kits for other low molecular weight molecules; i.e., mycotoxins.

The research is directed at developing immunoassay diagnostics for potential field-kit use that would more easily determine the ripening of pome fruit (e.g. fruit with internal seeds). The focus will be on developing enzyme-linked immuno system assays (ELISA's) which are specific and highly sensitive in detecting proteins associated with ripening. During Phase I polyclonal antibodies were produced against malic enzyme and post-initiation protein pAp5 and crude ELISA's were developed. In Phase II monoclonal antibodies will be produced to improve their specificity and sensitivity. Prototypes will be developed and validated. These ELISAs will be used to quantitate changes in specific proteins that occur in synchrony with or just prior to the onset of the ripening process. Efficacy tests will be performed with fruits from experimental controlled atmosphere (CA) storage which maintains fruits in the preclimacteric condition. Gene expression of these proteins also will be investigated by Michigan State University scientists. the diagnostics developed could have major economic benefits for fruit growers by minimizing losses incurred too early or too late picking of fruit.