Structure-specific hapten design for the screening of highly sensitive and specific monoclonal antibody to salbutamol

Salbutamol (SAL) is a short-acting beta 2-adrenergic receptor agonist (beta-agonist) forbidden in livestock production. To develop a more sensitive and specific detection method for SAL, a new hapten was synthesized, keeping the structure of benzene ring side of SAL while changing the tert-butyl group structure that SAL shares with other beta-agonists. Ten mice were immunized with a SAL hapten-keyhole limpet hemocyanin (KLH) conjugate, and cells were selected within a urine sample at the cell fusion stage. The best monoclonal antibody (Mab) had an IC50 value of 0.31 ng mL(-1) in PBST buffer and an IC50 value of 0.19 ng mL(-1) in a swine urine sample diluted five times (with a lower B-0 value), and had no cross-reactivity with clenbuterol and other beta-agonists. These results show that the hapten design was successful and that the Mab can potentially be used in highly sensitive immunoassays for specifically monitoring the illegal use of SAL in real samples.