Fragment-based hapten design and screening of a highly sensitive and specific monoclonal antibody for ractopamine

Abstract

Ractopamine (RCT) is a beta-adrenergic agonist and its use is forbidden in many countries. With the aim of more sensitive and specific detection of RCT, a RCT structure fragment-based hapten was synthesized by introducing a carboxyl group to octopamine through a reductive amination reaction with ethyl levulinate.Six mice were immunized with a hapten-KLH conjugate, and the best monoclonal antibody (Mab) with an IC 50 value of 0.34 ng mL À1 in PBST buffer and 0.41 ng mL À1 in a 10-times diluted swine urine sample was obtained, which had no cross reactivity with clenbuterol and salbutamol. These results demonstrated that a fragment of RCT as a hapten can mimic the antigenicity and specificity of RCT very well, and the Mab has potential applications in the specific monitoring of the illegal use of RCT in real samples.

Development of an ELISA and Immunochromatographic Assay for Tetracycline, Oxytetracycline, and Chlortetracycline Residues in Milk and Honey Based on the Class-Specific Monoclonal Antibody

“…The cell fusion procedure has been described elsewhere (Liu et al 2014b). Briefly, mice spleen was ground, yielding spleen cells.…”

Establishment of a monoclonal antibody-based indirect enzyme-linked immunosorbent assay for the detection of trimethoprim residues in milk, honey, and fish samples

“…Generally, cross-reactivity (CR) was measured to assess specificity. CR was calculated according to the following equation (Liu, Kuang, Peng, Wang, & Xu, 2014): CR% = (IC 50 value of TMP) (IC 50 value of related compound) × 100.…”

Development of an enzyme-linked immunosorbent assay (ELISA) for natamycin residues in foods based on a specific monoclonal antibody

“…Generally, the specicity was assessed by measuring the cross-reactivity (CR), which was calculated according to the following equation, 33 CR% ¼ (IC 50 value of Nata)/ (IC 50 value of related compound) Â 100. Generally, the specicity was assessed by measuring the cross-reactivity (CR), which was calculated according to the following equation, 33 CR% ¼ (IC 50 value of Nata)/ (IC 50 value of related compound) Â 100.…”