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采用光学生物传感器自动直接免疫分析法测定牛初乳粉、牛乳粉及含牛初乳产品的膳食补充剂中的牛免疫球蛋白G:协作研究

Determination of bovine immunoglobulin G in bovine colostrum powders, bovine milk powders, and dietary supplements containing bovine colostrum products by an automated direct immunoassay with optical biosensor: collaborative study.

作者信息

Gapper Leyton

机构信息

Fonterra Co-operative Group Ltd, Private Bag 11029, Palmerston North 4442, New Zealand.

出版信息

J AOAC Int. 2013 Sep-Oct;96(5):1026-32. doi: 10.5740/jaoacint.cs2012-26.

DOI:10.5740/jaoacint.cs2012-26
PMID:24282942
Abstract

Nine laboratories participated in an AOAC collaborative study to determine bovine immunoglobulin G (IgG) levels in selected dairy powders and dietary supplements by surface plasmon resonance (SPR) methodology. Each sample matrix was dissolved in buffer and suitably diluted to fit within the standard curve. The sample extract was injected over a surface functionalized with affinity-purified, polyclonal goat anti-bovine IgG (H+L) antibody; IgG was then detected. SPR detection was used for the direct immunoassay and quantification was made against a calibration curve prepared from bovine serum IgG. Between each standard and sample, the surface was regenerated using 10 mM glycine at pH 1.5. The samples analyzed included the likely matrixes for which the assay would find commercial use, namely, high- and low-protein-content colostrum powders, tablets containing colostrum powder, infant formula containing colostrum powder, and some IgG-containing dairy powders, i.e., milk protein isolate, whey protein concentrate, and skim milk powder. Each laboratory provided data for the study and assayed blind duplicates of seven materials. Due to gross outliers in the majority of results from one laboratory, the data from eight laboratories were used for the statistical analysis. The repeatability RSD (RSDr) values ranged from 3.2 to 7.3%, and the reproducibility RSDR values from 13.0 to 22.6%.

摘要

九个实验室参与了一项美国官方分析化学师协会(AOAC)的协作研究,旨在通过表面等离子体共振(SPR)方法测定特定奶粉和膳食补充剂中的牛免疫球蛋白G(IgG)水平。将每个样品基质溶解在缓冲液中,并进行适当稀释以符合标准曲线范围。将样品提取物注入用亲和纯化的多克隆山羊抗牛IgG(H + L)抗体功能化的表面;然后检测IgG。SPR检测用于直接免疫测定,并根据由牛血清IgG制备的校准曲线进行定量。在每个标准品和样品之间,使用pH 1.5的10 mM甘氨酸再生表面。分析的样品包括该检测方法可能用于商业用途的基质,即高蛋白和低蛋白含量的初乳粉、含初乳粉的片剂、含初乳粉的婴儿配方奶粉以及一些含IgG的奶粉,即乳清蛋白分离物、乳清蛋白浓缩物和脱脂奶粉。每个实验室提供了该研究的数据,并对七种材料进行了盲法重复检测。由于一个实验室的大多数结果中存在严重异常值,因此使用了八个实验室的数据进行统计分析。重复性相对标准偏差(RSDr)值范围为3.2%至7.3%,再现性相对标准偏差(RSDR)值范围为13.0%至22.6%。

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