A specific monoclonal antibody for chlortetracycline detection in milk and honey samples based on ELISA

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 IC 50 of 0.33 ng/ml and a detection limit of 0.1 ng/ml. The cross reactivity of this antibody with structurally related and nonrelated 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.

Development of a colloidal gold strip assay for the detection of total homocysteine in serum samples

“…The antibody was purified from the collected ascites by protein G affinity chromatography according to the manufacturer's instructions. 25 The purified antibody was dialyzed against PBS for 2 days at 4 °C and then stored at −20 °C.…”

Fluorescent strip sensor for rapid and ultrasensitive determination of fluoroquinolones in fish and milk

“…The mouse with the highest OD 450 nm values and inhibition ratio was selected to be sacrificed for the preparation of spleen cells, and fusion of splenocytes and Sp2/ 0 myeloma cells was performed using PEG 4000 in a sterile environment. [35][36][37] The fused cells were distributed into 96-well plates and cultured in HAT medium for the first five days to inhibit the growth of unfused myeloma cells or hybrid cells formed by conspecific cell fusion, and then transferred into the normal HT medium for subsequent culture. After 7 days of cell fusion, ic-ELISA was used to test the cell supernatant in 96-well plates.…”

Sensitive immunochromatographic assay for the detection of the dimethachlone fungicide in tomatoes and lettuces

“…17 Then, the mAbs were collected by in vivo induction of ascites in mice and purified by caprylic acid/ammonium sulphate precipitation (CA-SA). [18][19][20] The purified anti-DMT mAbs were identified by ELISA and the titer, affinity constant, and specificity were determined as previously reported. [21][22][23]…”