DEVELOPMENT OF MONOCLONAL-ANTIBODY FOR ROCK SHRIMP IDENTIFICATION USING ENZYME-LINKED-IMMUNOSORBENT-ASSAY

Rock shrimp specific protein M, determined from a comparison of the SDS-PAGE protein banding pattern of rock shrimp with those of 23 other seafood and meat samples, was isolated from 12% polyacrylamide gels and used to develop monoclonal antibodies (McAb). The highly specific McAb (4H2-10D3) recognized the antigenic determinant(s) present on the 17.7-kDa component of protein M in both the native and heat-denatured rock shrimp extract. Specificity of the McAb was confirmed by testing against pink, white, and rock shrimp protein extracts using Western blotting technique and also against 23 commonly consumed seafood and meat samples by immunodot blotting and ELISA. The McAb was also applied in ELISA under optimized test conditions to detect the presence of rock shrimp in 26 various seafood and meat mixture samples. The McAb correctly detected rock shrimp in all these samples. Heat treatment of rock shrimp extract enhanced the reactivity with this McAb. Furthermore, the presence of rock shrimp as low as 4.3 ng on the average in sample mixtures containing various seafood or meat samples could be detected. © 1990, American Chemical Society. All rights reserved.