免疫磁珠快速法：检测牛奶中β-酪蛋白的高效新途径

169 浏览量更新于2024-08-26 收藏 625KB PDF 举报

本研究报道了一种创新的快速检测技术——基于免疫磁珠的酶联免疫吸附试验（IMBs-ELISA），用于在牛乳中精确检测β-酪蛋白。免疫磁珠（IMB）在该实验中扮演了关键角色，作为固相支持，其独特的磁性特性使得样本处理更加高效。研究者利用抗β-酪蛋白的单克隆抗体（McAb）作为捕获探针，将其固定在IMB上，形成固相抗体复合物。随后，辣根过氧化物酶（HRP）标记的抗β-酪蛋白的多克隆抗体（PcAb）作为检测探针，通过特异性结合被捕获的β-酪蛋白进行信号放大。与传统ELISA相比，IMBs-ELISA显著减少了反应步骤，每个步骤仅需10分钟或更短，提高了整体检测速度。这一设计采用了夹心结构，即先用McAb捕捉目标蛋白质，再用PcAb进行二次识别，确保了检测的敏感性和特异性。实验结果显示，在线性范围内（2-128μg/mL）准确地测定了牛乳中的β-酪蛋白含量，其结果与公认的Kjejdahl方法相符，表明该方法具有良好的重现性和准确性。这项工作在牛乳质量控制、食品安全监测以及β-酪蛋白相关研究中具有重要意义，因为它不仅提供了快速检测手段，还可能有助于提高乳制品行业的生产效率和产品质量控制标准。同时，免疫磁珠技术的运用展示了其在生物传感器和快速检测领域广阔的应用前景。

Analytical Methods

A rapid immunomagnetic beads-based immunoassay for the detection

of b-casein in bovine milk

F. Song

, Y. Zhou

⇑

, Y.S. Li

, X.M. Meng

, X.Y. Meng

, J.Q. Liu

, S.Y. Lu

, H.L. Ren

,P.Hu

, Z.S. Liu

Y.Y. Zhang

, J.H. Zhang

Key Laboratory of Zoonosis, Ministry of Education, Institute of Zoonosis/College of Veterinary Medicine, Jilin University, Changchun 130062, PR China

Grain and Oil Food Processing Key Laboratory of Jilin Province, Jilin Business and Technology College, Changchun 130062, PR China

Production Quality Test Institute of Jilin Province, Changchun 130022, PR China

article info

Article history:

Received 19 October 2012

Received in revised form 22 September 2013

Accepted 26 February 2014

Available online 12 March 2014

Keywords:

Immunomagnetic beads

Sandwich structure

b-Casein

Enzyme-linked immunosorbent assay

Detection

abstract

An immunomagnetic beads-based enzyme-linked immunosorbent assay (IMBs-ELISA) was developed for

the detection of b-casein in bovine milk. Immunomagnetic beads (IMBs) were employed as the solid

phase. The anti-b-casein monoclonal antibody (McAb) bound to IMBs was used as capture probe and

an anti-b-casein polyclonal antibody (PcAb), labelled with horseradish peroxidase (HRP), was employed

as detector probe. Three reaction and two washing steps were needed. Each reaction needed 10 min or

less, which signiﬁcantly shortened detection compared with classic sandwich ELISA. b-Casein in bovine

milk w as detected across a linear range (2–128

gmL

1

). Application results were in accordance with

the Kjejdahl method, which suggests the IMBs-ELISA is rapid and reliable for the detection of b-casein

in bovine milk.

1. Introduction

Bovine milk is a dairy product with high nutritional value that is

popular amongst consumer of all ages. It contains protein, fat, car-

bohydrate, minerals, vitamin and water. In recent years, the con-

sumption of dairy products has rapidly increased (Bashir, 2011).

However, milk and milk product have become the target of adulte-

ration (fraudulent incorporation of less costly ingredients), which

is a signiﬁcant problem for the dairy industry in many countries

in the world and consumers, for example, the pet-food contamina-

tion by melamine adulterant in America in 2007 (Dobson et al.,

2008) and the melamine-tainted-milk powder event in China in

September 2008 (Chen, 2009). The common adulterants also

include urea, nitrates, alum, and soya-bean meal, besides

melamine (Attia, Bakir, Abdel-aziz, & Abdel-mottaleb, 2011), which

are all nitrogen-containing materials. There is no distinction

between protein nitrogen and non-protein nitrogen when the

adulterate milk is tested using the standard Kjeldahl method,

which only measures total nitrogen not nitrogen source/types,

and gives a false impression of levels of milk protein content. It

may be harmful to human health and is unethical. There is a need

to develop a rapid and reliable method not only for such safety

problems but also for quality in cases of adulteration with other

milks (Chen, 2009). Milk quality is generally evaluated by

determining total proteins, and it contains whey proteins (20%)

and caseins (80%) (Muller-Renaud, Dupont, & Dulieu, 2004). There

are four kinds of caseins in bovine milk, namely

S1- (37%),

S2- (10%), b- (37%), and

-caseins (10%), respectively (Johansson

et al., 2009). Among these, b-casein is the major indigenous. It con-

sistently makes up 35–45% total casein content (Colin, Laurent, &

Vignon, 1992; Remeuf, Lenoir, & Duby, 1989; Song, Xue, & Han,

2011). Thus, the quantity of b-casein in bovine milk could be used

as an index to evaluate its quality and detect dairy adulteration.

Various analytical techniques have been proposed for milk authen-

tication including optical immunosensor (Muller-Renaud et al.,

2004), single frequency electrical conductance measurements

(Mabrook & Petty, 2003), isoelectric focusing (Kim & Jimenez-Flo-

res, 1994; Rodríguez, Ortiz, Sarabia, & Gredilla, 2010), capillary

electrophoresis (Miralles, Ramos, & Amigo, 2000; Recio, Amigo, &

Lopez-Fandino, 1997), hydrophobic interaction chromatography

(Bramanti, Sortino, Onor, Beni, & Raspi, 2003) and enzyme-linked

immunosorbent assay (ELISA) (Hurley, Coleman, Ireland, &

Williams, 2006). However, most of these methods require sophis-

ticated, technical expertise.

Immunomagnetic beads (IMBs) have more surface area than a

ﬂat solid phase, permitting more ‘active molecules’ to be

immobilised on the surface and enhancing sensitivity (Soh et al.,

http://dx.doi.org/10.1016/j.foodchem.2014.02.150

⇑

Corresponding author. Tel.: +86 0431 87835734; fax: +86 13634318992.

E-mail address: zhouyu69@sina.com (Y. Zhou).

Food Chemistry 158 (2014) 445–448

Contents lists available at ScienceDirect

Food Chemistry

journal homepage: www.elsevier.com/locate/foodchem

下载后可阅读完整内容，剩余3页未读，立即下载

weixin_38605801

粉丝: 10
资源: 984

免疫磁珠快速法：检测牛奶中β-酪蛋白的高效新途径

行业文档-设计装置-一种β-酪蛋白单克隆抗体的制备方法和β-酪蛋白胶体金检测试纸条及其制备方法.zip

基于金纳米粒子聚集的比色测定法用于牛乳样品中的β-酪蛋白检测

基于双抗体的免疫测定法，用于检测牛乳样品中的β-酪蛋白。

牛乳中α-酪蛋白完全抗原的合成及多克隆抗体的制备

行业文档-设计装置-一种β-酪蛋白胶体金检测试纸条.zip

母乳喂养和奶瓶喂养婴儿的牛 β-酪蛋白抗体：它们与 1 型糖尿病的相关性

四种二肽对奶牛乳腺上皮细胞增殖及β-酪蛋白分泌的影响 (2013年)

消除乳清蛋白中β-乳球蛋白致敏性的研究进展 (2009年)

行业分类-设备装置-αs1-酪蛋白和κ-酪蛋白特异性抗体及试剂盒和试纸条.zip

β-环糊精包埋技术及研究进展 (2010年)

最新资源