School of Food and Biological Engineering, Jiangsu University, Zhenjiang, China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2011 Jul 15;879(22):2149-54. doi: 10.1016/j.jchromb.2011.05.053. Epub 2011 Jun 12.
Methandrostenolone (MA) is a steroid used as veterinary medicine on stockbreeding to promote animal growth. The use of MA has been strictly regulated because of its harmful effect on consumers. This paper describes the production of polyclonal antibody (pAb) against MA, the preparation of immunoaffinity column (IAC) and its potential application to the selective extraction of MA residues from animal tissue and feed samples. The produced pAb exhibited good sensitivity to MA with an IC(50) value of 5.6 ng/mL. The cross-reactivity values of the antibody with MA structurally related compounds of testosterone propionate (TP) and trenbolone (TR) were lower than 0.6%. By coupling the produced antibody with CNBr-activated Sepharose 4B, an IAC was prepared. 2% methanol and 80% methanol were selected as loading and eluting solution by optimization. The maximum capacity of the column for MA was approximately 334 ng/mL gel. The average recovery of 20, 40 and 60 ng/mL MA standard solutions from IACs was 97.9% with the relative standard deviation (RSD) among columns of 6.7%. After 3 times of repeated usage, the column capacity and recovery rate still remained 82.0% and 92.6% respectively. The IACs were then challenged with MA-fortified animal tissue and feed samples, recoveries of MA were found to be in the range of 83.5-99.7%.
美雄酮（MA）是一种兽用类固醇，用于畜牧业以促进动物生长。由于对消费者有有害影响，MA 的使用受到严格监管。本文介绍了针对 MA 的多克隆抗体（pAb）的生产、免疫亲和柱（IAC）的制备及其在从动物组织和饲料样品中选择性提取 MA 残留方面的潜在应用。所产生的 pAb 对 MA 具有良好的灵敏度，IC 50 值为 5.6 ng/mL。该抗体与 MA 结构相关化合物丙酸睾酮（TP）和群勃龙（TR）的交叉反应值均低于 0.6%。通过将产生的抗体与 CNBr 活化的 Sepharose 4B 偶联，制备了 IAC。通过优化，选择 2%甲醇和 80%甲醇作为上样和洗脱溶液。该柱对 MA 的最大容量约为 334 ng/mL 凝胶。从 IAC 中提取 20、40 和 60 ng/mL MA 标准溶液的平均回收率分别为 97.9%，柱间相对标准偏差（RSD）为 6.7%。重复使用 3 次后，柱容量和回收率仍分别保持在 82.0%和 92.6%。然后用 MA 强化的动物组织和饲料样品对 IAC 进行了挑战，发现 MA 的回收率在 83.5-99.7%范围内。
