Production and Characterization of Anti-idiotype and anti-anti-idiotype Antibodies from a Monoclonal Antibody against Aflatoxin

Anti‐idiotype and anti‐anti‐idiotype antibodies generated from polyclonal antibodies against aflatoxin b1

“…The indirect ELISA showed that pAb2 bound specifically to the originated pAbl against AFBi, but not to immunoglobulins from other irrelevant monoclonal antibodies or to normal mouse serum. In addition, the original pAbl used in this present study did not show significant interaction with anti-id antibodies generated from mice that were either immunized with mAbl against AFB 3 -HS-BSA or with the mAbl conjugated to KLH (Hsu & Chu, 1994). Data obtained from both direct and indirect ELIS As indicated that the pAb2 effectively inhibited the binding of AFBi to original idiotype antibodies.…”

“…*AFB 3 -mAbl, AFB 3 -pAbl and AFB 3 -pAb3 are monoclonal antibodies against AFB 3 -HS-BSA conjugate (Hefle & Chu, 1990), rabbit polyclonal antibodies against AFB 3 -HS-BSA conjugate (Zhang & Chu, 1989;Hsu & Chu, 1994) and mouse polyclonal anti-anti-id antibodies against AFB3-HS-BSA (Hsu & Chu, 1994) respectively.…”

“…The affinity gel was prepared under conditions similar to those described before (Hsu & Chu, 1994). For purification, lml of pAbl (containing 20 mg of protein) was applied to the column, recycled three times and then left to stand for 30 min at room temperature.…”

“…To eliminate non-related and pre-existing non-specific antibodies, a CH-Sepharose 4B column armed with purified pAbl was used. The column Downloaded by [New York University] at 14:34 12 April 2015 was prepared by conjugation with 10 mg of purified pAbl to 0.5 g of cyanogen bromideactivated CH-Sepharose 4B according to the manufacturer's instructions as detailed before (Hsu & Chu, 1994).…”

“…Some anti-id polyclonal antibodies (Ab2) have been shown to mimic effectively the biological functions of the hapten (Nisonoff, 1991). Recently, anti-id antibodies against an anti-aflatoxin B 3 (AFB 3 )-hemisuccinate (HS)-bovine serum albumin (BSA) monoclonal antibody from rabbits have been obtained and characterized (Hsu & Chu, 1994). In this study, rabbit polyclonal antibodies against AFBi-carboxymethyloxime (CMO)-BSA conjugate having high sensitivity and specificity to AFBi in enzyme-linked immunosorbent assay (ELISA) and radioimmunoassay (RIA) systems were chosen as the idiotype antibody (pAbl) for generating pAb2 from mice.…”