Preparation of an immunoaffinity column and its application in sample cleanup for methandrostenolone residues detection

Wang, Yun; Xu, Yan; Zhang, Xun; Wang, Enlan; Dong, Ying

doi:10.1016/j.jchromb.2011.05.053

Cited by 9 publications

(3 citation statements)

References 30 publications

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“…Therefore, the maximum binding capacity of this IAC for MA was calculated as [5 )200 (completely retained) '190 (partially retained)]/0.25 04760 ng/mL gel. Because of the better homogeneity of mAb compared with that of pAb, the column capacity obtained in this study is much higher than that of our previous reported pAb-based IAC (Wang et al, 2011a).…”

Section: Characterisation Of the Iacmentioning

confidence: 62%

“…Owing to the advantages of high specificity, simple operation and therefore low cost per sample, immunoaffinity chromatography has been widely used in sample pretreatment for residue analysis of mycotoxins (Klinglmayr, Nöbauer, Razzazi-Fazeli, & Cichna-Markl, 2010;Tavčar-Kalcher, Vrtač, Pestevšek, & Vengušt, 2007;), veterinary drugs (Wang & Shen, 2007;Zhang et al, 2011), pesticides (Chen, Wang, Wang, & Tang, 2009;Wang et al, 2011) and heavy metals (Abe et al, 2011). Our previous work has described an immunoaffinity extraction of MA from food and feed samples using a polyclonal antibody (pAb) against MA (Wang, Xu, Zhang, Wang, & Dong, 2011a). Compared with pAb, monoclonal antibody (mAb) can be generated as a constant and renewable resource with better homogeneity and consistency (Lipman, Jackson, Trudel, & Weis-Garcia, 2005;Stanker, Merrill, Scotcher, & Cheng, 2008).…”

Section: Development and Characterisation Of An Immunoaffinity Columnmentioning

confidence: 99%

“…The preparation of haptens and conjugates and the generation of anti-MA mAbs were conducted according to our previous reports (Wang et al, 2011a;Wang, Xu, Zhang, Wang, & Dong, 2011b). In brief, a carboxyl group was introduced into MA by oximate reaction firstly and then the synthetic hapten MA (Figure 1) was covalently coupled with keyhole limpet haemocyanin (KLH) and bovine serum albumin (BSA), respectively, using the modified active ester method to obtain immunogens and coating antigens.…”

Section: Preparation and Evaluation Of Monoclonal Anti-ma Antibodiesmentioning

confidence: 99%

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Development and characterisation of an immunoaffinity column for the selective extraction of methandrostenolone from food and feed samples

Wang

Zhang

et al. 2013

Food and Agricultural Immunology

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2013)Development and characterisation of an immunoaffinity column for the selective extraction of methandrostenolone from food and feed samples, Food and Agricultural Immunology, 24:2, 231-240, Methandrostenolone (MA) is used as veterinary medicine for promoting foodproducing animal growth. Accumulation of MA in a human body would cause multiple side effects. In this study, an anti-MA monoclonal antibody (mAb)-based immunoaffinity column (IAC) was prepared and its application to the selective extraction of MA from actual samples was evaluated. The employed antibody exhibited good sensitivity and specificity towards MA with an IC 50 value of 7.89 ng/mL and less than 1% cross-reactivity towards structurally related compounds. By coupling CNBr-activated Sepharose with the antibody, an IAC was prepared. Two percent methanol and 70% methanol were selected, respectively, as loading and eluting solution by optimisation. The maximum binding capacity of the column for MA reached 4760 ng/mL gel. The average recovery of 50, 250 and 500 ng MA standards from IACs was 88.6% with a relative standard deviation (RSD) among columns of 8.5%. After three times usage, the binding capacity and recovery still remained 93.4% and 81.1%, respectively. To further verify the effect of matrices on the extraction efficiency, the IACs were challenged with MA-fortified animal tissue and feed samples and the recovery were found to be in the range of 78.7%Á95.1%.

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Section: Characterisation Of the Iacmentioning

confidence: 62%