29G-15 |
Optimization of the immobilization of polyclonal and monoclonal antibodies to detect Salmonella spp. and Campylobacter spp. |
H. NASRI1, I. Goktepe2, and M. Ahmedna2. (1) Food Science and Nutrition, North Carolina A&T State University, 171 Carver Hall, Greensboro, NC 27411, (2) Dept. of Human Environment & Family Science, North Carolina A&T State Univ., 161 Carver Hall, Greensboro, NC 27411 Foodborne salmonellossis and compylobacteriosis continue to be significant problems for the food industry, and public health. Early detection of Salmonella and Campylobacter is of the utmost importance in the detection, recognition, and effective prevention of an outbreak. Enzyme-linked immunosorbent assays (ELISAs) are widely used for monitoring of bacteria including bacteria, viruses, and other microorganisms due to their high reproducibility and possibility for simultaneously conducting a large number of assays. However, several researchers pointed out that conventional ELISA procedures carried out by immobilizing antigen or antibody on a polystyrene microtiter plate through adsorption suffer from inconsistent ELISA values in different wells and plates, nonreproducible results due to detachment of biomolecules during washing and lower sensitivity. The most critical step to increase detection system in ELISA is the binding of the capture antibodies to the solid surface. The aim of this study is to compare the capacity of monoclonal and polyclonal antibodies to bind to two types of adsorption surfaces. The two surfaces used were a polystyrene surface, which allows a passive adsorption of the antibodies and an aminated polystyrene surface, which allows a covalent adsorption when used with a crosslinker like BS3. Aminated ELISA plates are solid surfaces modified to yield amino groups for covalent binding of antibodies. The use of the aminated membranes resulted in 5 to 10% improvement over the standard polystyrene membranes. Furthermore, the current study shows that the use of polyclonal antibodies as capture antibodies resulted in higher ELISA absorbance values than monoclonal when used on aminated surfaces. Polyclonal were more efficient than monoclonal antibodies in detecting Salmonella spp. and Campylobacter spp. when used in combination with a polyclonal conjugate especially when used on aminated surfaces.
Session 29G, Food Microbiology: General
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