A specific monoclonal antibody for chlortetracycline detection in milk and honey samples based on ELISA
Panicha Poungmalai, Anumart Buakeaw, Songchan Puthong, Nanthika Khongchareonporn
Abstract
In this study, a monoclonal antibody (mAb) against chlortetracycline was prepared by immunisation using chlortetracycline conjugated to bovine serum albumin (CTC-BSA) and cell fusion. One monoclonal antibody (1-2B) against chlortetracycline was obtained with an IC50 of 0.33 ng/ml and a detection limit of 0.1 ng/ml. The cross reactivity of this antibody with structurally related and non-related compounds was less than 0.1%, demonstrating that the obtained mAb provided high specificity. Under optimal conditions for ic-ELISA, a linearity was obtained between 0.06 and 3.9 ng/ml of chlortetracycline. The quantification of CTC-spiked milk and honey samples was also performed, demonstrating recovery rates ranging from 95.2% to 105.6% and the coefficient of variation below 10% for both intra- and inter-variation assays. Based on the sensitivity and specificity measurement, mAb 1-2B could feasibility be used as a screening tool for CTC residue detection in milk and honey samples.