Papers by Author: Jin Qing Jiang

Abstract: This paper reports an indirect competitive enzyme-linked immunosorbent assay (icELISA) using polyclonal antibody (pAb) for estradiol (E2) residues. After derivation, E2 haptens were conjugated to bovine serum albumin (BSA) and ovalbumin (OVA) through 1-Ethyl-3-(3-dimethylaminopropy) carbodiimide (EDC) method, and New Zealand white rabbits were immunized to produce anti-E2 pAb. The conjugation ratio of E2-BSA was proved to be 18.6:1 by an UV absorbance method. Based on the square matrix titration, an icELISA standard curve was developed. The dynamic range was from 0.16 to 128 ng/mL, with LOD and IC50 value of 0.08 ng/mL and 3.76 ng/mL, respectively. Except for a little cross-reactivity (16.2%) to estrone, this assay showed negligible cross-reactivity to other analogues tested. The results suggest that the produced anti-E2 pAb could be used to develop an icELISA method for the determination of E2 residues in animal-originally products.

Abstract: This study aimed to develop a monoclonal antibody based icELISA method for Ractopamine (Rac) residue. For this purpose, mixed anhydride method was employed to synthesize the immunogen of Rac-BSA and 1, 4-butanediol diglycidyl ether was used to prepare the coating antigen of Rac-OVA, thus pursue the heterologous sensitivity. Through cell fusion technology, four Hybridoma named R1-B5, R2-B3, R2-C6, and R4-C8 were screened out, and the Kas of all mAbs were between 2.7 and 4.8×109 L/mol. Based on the R1-B5 mAb, a heterologous icELISA standard curve was developed. The working range was from 0.013 to 33.7 ng/mL, with LOD and IC50 value of 0.007 ng/mL and 0.67 ng/mL, respectively. Therefore, this icELISA can be used for detecting Rac residue in animal products.

Abstract: Through cell fusion technology, five hybridoma lines of sarafloxacin (SAR), named S1-B2, S2-C6, S2-E7, S3-C5, and S3-E5, were identified and their corresponding mAbs were of the IgG1 isotype with a k light chain. The Kaffs of all mAbs were between 2.8 and 4.6×109 L/mol. The titers and IC50 values of purified ascite fluids were in the range of 0.512–2.56×106 and 0.32–0.48 ng/mL, respectively. The performances of S1-B2 and S2-C6 were more consistent in the stability experiments. Based on the S1-B2 hybridoma, an icELISA method was developed. The dynamic range was from 0.004 to 18 ng/mL, with a detection limit for the assay and IC50 values of 0.002 and 0.32 ng/mL, respectively. Therefore, the establishment of these hybridomas may provide an alternative method for the detection of SAR residues in food-original animals.

Abstract: A rapid indirect competitive ELISA (icELISA) for the determination of enrofloxacin (ENR) residue has been developed. EDC method was employed to synthesize the artificial antigen of ENR-BSA, and anti-serum produced from rabbits was selected. Based on the square matrix titration, an icELISA method was developed with the polyclonal antibody. The Linear range was from 0.006 to 31.5 ng/mL, with LOD and IC50 values of 0.003 ng/mL and 0.45 ng/mL, respectively. Except for a high cross-reactivity (105.2%) to ciprofloxacin, negligible cross-reactivity to other compounds was observed. The recoveries of ENR were in the range of 98.3-127.5%, 85.7-112.5% and 97.4-103.8% for chicken muscle, liver and kidney, respectively. Therefore, this assay has the potential to be incorporated into a quantitative monitoring program for the rapid screening of ENR residue in food.

Abstract: This article aimed to optimize a polyclonal antibody based indirect competitive ELISA (icELISA) and compare between the icELSIA method and the LC-MS technology for the determination of enrofloxacin (ENR) residue. Based on the square matrix titration, linear range of the icELISA was from 0.006 to 31.5 ng/mL, with LOD and IC50 values of 0.003 ng/mL and 0.45 ng/mL, respectively. After optimization, 0.03 mol/L of HCl was used in the assay buffer and this ELISA system can tolerate acetonitrile not higher than 10%. The recoveries of ENR were in the range of 98.3-127.5%, 85.7-112.5% and 97.4-103.8% for chicken muscle, liver and kidney. The correlation coefficients (R2) of the ELISA and LC-MS data were 0.9472 in muscle, 0.9843 in liver, and 0.9382 in kidney, respectively.