Preparation of Anti-Tetracycline Antibodies and Development of an Indirect Heterologous Competitive Enzyme-Linked Immunosorbent Assay to Detect Residues of Tetracycline in Milk
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- 作者：Yulan Zhang ; Shengxin Lu ; Wei Liu ; Chengbiao Zhao ; Rimo Xi
- 刊名：Journal of Agricultural and Food Chemistry
- 出版年：2007
- 出版时间：January 24, 2007
- 年：2007
- 卷：55
- 期：2
- 页码：211 - 218
- 全文大小：156K
- 年卷期：v.55,no.2(January 24, 2007)
- ISSN：1520-5118
文摘
Tetracycline (TC) is a broad-spectrum antibiotic used increasingly in animal husbandry to treat diseasesor to promote growth as feed additives. To avoid using labor-intensive instrumental methods to detectresidues of TC in food and food products, a simple and convenient indirect heterologous competitiveenzyme-linked immunosorbent assay (ELISA) method for TC was developed using polyclonal antibodyprepared in this study. Three new immunogens, TC-o-tolidine-bovine serum albumin (BSA), TC-4-aminobenzoic acid-cationized BSA (cBSA), and TC-1,1'-carbonyldiimidazole-cBSA, were synthesized in this research to develop anti-TC antibodies. All antibodies raised in rabbits and coatingantigens synthesized were screened and characterized using homologous and heterologous ELISAformats to select the best combination. An optimized ELISA gave an IC50 value of 3.92 
g/mL towardTC in PBS buffer. The specificity of the assay was studied by measuring cross-reactivity of the antibodywith the structurally closely related compounds of chlortetracycline (112%) and oxytetracycline (<2%).The recovery rates from the TC-fortified raw milk samples were in the range of 74-116%, while theintra- and interassay coefficients of variation were <14.5 and <25.0, respectively.
g/mL towardTC in PBS buffer. The specificity of the assay was studied by measuring cross-reactivity of the antibodywith the structurally closely related compounds of chlortetracycline (112%) and oxytetracycline (<2%).The recovery rates from the TC-fortified raw milk samples were in the range of 74-116%, while theintra- and interassay coefficients of variation were <14.5 and <25.0, respectively.