Preparation and Selection of Polyclonal Antibodies against Malachite Green
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Abstract:
Three haptens for malachite green were designed and synthesized. Then the haptens were separately coupled to keyhole limpet hemocyanin (KLH) as immunogens and coupled to ovalbumin (OVA) as coating proteins by using active ester method. New Zealand white rabbits were immunized with the three immunogens, and all produced high titer antisera antibodies angainst malachite green. All the antibodies and the coating proteins were screened and then the optimal antibody-coated antigen combination was obtained. The selected polyclonal antiserum was purified by Sepharose FF-Proteins A affinity chromatography. The titer and characteristic were determined by indirect ELISA. The results showed that the antibody titer and IC50 reached 1:12000, and 0.65 ng/mL, respectively. And the antibody had good specificity. The cross-reactivity was less than 10% for leucomalachite green and leucocrystal violet but the antibody exhibited significant cross reactivity with crystal violet (18.73%). The study provided basis for rapid detecting malachite green residues in food by immune analysis.
