Indyk Harvey E, Filonzi Enrico L
NZMP-Fonterra, PO Box 7, Waitoa, New Zealand.
J AOAC Int. 2003 Mar-Apr;86(2):386-93.
An automated biosensor surface-plasmon resonance-based assay was developed for the determination of immunoglobulin G (IgG) in bovine milk and colostrum with either goat or rabbit antibovine IgG or protein G used as detecting molecule. The method is configured as a direct and nonlabeled immunoassay, with quantitation against an authentic IgG calibrant. Whole colostrum or milk is prepared for analysis by dilution into buffer. Analysis conditions, including ligand immobilization, flowrate, contact time, and regeneration, were optimized, and nonspecific binding was evaluated. Performance parameters included working range of 15-10 000 ng/mL, method detection limit of 0.08 mg/mL, overall instrument response reproducibility relative standard deviation (RSD(R)) of 0.47%, mean between-run RSD(R) of 10.5% for colostrum, and surface stability over 200 analyses. The proposed method was compared with independent alternative methods. The technique was applied to the measurement of IgG content during early lactation transition from colostrum to milk, as well as in consumer milk, colostrum, and hyperimmune milk powders.
开发了一种基于自动生物传感器表面等离子体共振的检测方法，用于测定牛奶和初乳中的免疫球蛋白G（IgG），使用山羊或兔抗牛IgG或蛋白G作为检测分子。该方法配置为直接非标记免疫测定法，采用真实IgG校准物进行定量。将全初乳或牛奶稀释到缓冲液中制备用于分析的样品。对分析条件进行了优化，包括配体固定、流速、接触时间和再生，并评估了非特异性结合。性能参数包括工作范围为15 - 10000 ng/mL、方法检测限为0.08 mg/mL、仪器整体响应重现性相对标准偏差（RSD(R)）为0.47%、初乳批间平均RSD(R)为10.5%以及超过200次分析的表面稳定性。将所提出的方法与独立的替代方法进行了比较。该技术应用于早期泌乳从初乳向牛奶过渡期间以及市售牛奶、初乳和高免疫奶粉中IgG含量的测量。
