Screening of penicillin residues in milk by a surface plasmon resonance-based biosensor assay: comparison of chemical and enzymatic sample pre-treatment

A biosensor-based immunoassay for the screening of penicillin residues in m ilk was developed. The BIAcore biosensor which was employed was based on su rface plasmon resonance detection. One commercial antibody against ampicill in which had much higher affinity for open beta-lactam ring than for closed ring was chosen. Two different pre-treatments were tested prior to the bio sensor assay to open the ring in order to increase the assay sensitivity. T hese two ways were enzymatic (penicillinase) and chemical pre-treatment of the samples. Limits of detection for ampicillin in milk were 33 and 12.5 mu g l(-1) after enzymatic and chemical pre-treatment, respectively. There wer e no cross-reactions with cephalosporins and other families of antibiotics or proteins in the buffer and in milk after both the pre-treatments. Percen tages of cross-reactivity with nine tested penicillins were often clearly h igher in milk than in buffer. Moreover, a lack of sensitivity after penicil linase pre-treatment was observed. Conversely, penicillins G and penicillin s M could be detected at or below their respective MRLs (4 and 30 mug l(-1) in milk) and ampicillin and amoxicillin could be detected only at about th ree MRLs after chemical pre-treatment. However, the enzymatic pre-treatment was much easier to perform and led to more stable results. (C) 2001 Elsevi er Science B.V. All rights reserved.