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An aptamer affinity column for purification and enrichment of lactoferrin in milk

文献类型： 外文期刊

作者： Wang, Nan 1 ; Jiang, Xiaoqian 1 ; Xu, Xiaoling 3 ; Liu, Yan 3 ; Liu, Longfei 4 ; Lu, Anxiang 1 ; Lu, Jinghua 1 ; Luan, Yu 1 ;

作者机构： 1.Jinzhou Med Univ, Coll Pharm, Jinzhou 121001, Peoples R China

2.Beijing Res Ctr Agr Stand & Testing Agr Product Q, Dept Agr, Safety Risk Assessment Lab, Beijing Municipal Key Lab Agr Environm Monitoring, Beijing 100097, Peoples R China

3.Inst Anim Husb & Vet Med, Lab Anim Prod, Beijing 100097, Peoples R China

4.Beijing Biotai Co Ltd, Beijing 100097, Peoples R China

关键词： Aptamer; Lactoferrin; Aptamer affinity column; Milk

期刊名称：JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES （ 影响因子：3.205； 五年影响因子：3.068 ）

ISSN： 1570-0232

年卷期： 2021 年 1178 卷

页码：

收录情况： SCI

摘要： As an active glycoprotein with high nutritional value, lactoferrin is widely used in food and medical treatment. Therefore, it is very important to establish an accurate and efficient detection method for lactoferrin. At present, the detection of lactoferrin in milk faces many challenges, such as low separation degree and poor parallelism. To address this issue, we developed an aptamer affinity column (AAC) for purification and enrichment of lactoferrin in milk. The column was prepared by covalent conjugation of an amino-modified aptamer with NHS-activated Sepharose. The washing buffer type (0.01 mol/L phosphate buffer) and volume (10 mL) and the sodium chloride concentration (1 mol/L) in the elution buffer were optimized for the AAC method. The performance of the AAC was then evaluated in terms of the column capacity, specificity, stability, and reusability. The column capacity was 500 +/- 13.7 mu g and the column could be reused up to ten times with a large loss in performance. The AAC method combined with high-performance liquid chromatography gave excellent linearity over a wide range, good sensitivity with a limit of detection of 3 mu g/mL, and acceptable recoveries for different concentrations of lactoferrin spiked in real raw milk samples from cattle. Finally, the AAC was successfully applied to analyze lactoferrin in milk. This method could be applied to routine analysis of samples for lactoferrin in testing laboratories and dairy factories.

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