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[Optimizing the Detection of Bovine Milk β-Lactoglobulin With Protein Chip]

[Optimizing the Detection of Bovine Milk β-Lactoglobulin With Protein Chip]

Xinxin Ma, Jiyong Yin, Jing Sun, Jian Huang, Wei Piao, Jin Li, Di Chen, Xiaolin Yuan, Junsheng Huo

Wei Sheng Yan Jiu. 2017 Jan;46(1):78-83.

PMID: 29903156

Abstract:

Objective:
To optimize the conditions of protein chip assay for bovine milk β-Lactoglobulin( β-Lg).
Methods:
A microarrayer was used for printing anti-β-Lg as antibody I on each 3-dimensional-slide, another antis β-Lg antibody was used as detection antibody II and goat antibody coupled to Cy3 was used as antibody III. The standard β-Lg was detected by double antibody sandwich technique.
Results:
Mouse monoclonal β-Lg antibody66# was chosen as the probe and contact printing as the printing method. The range between 42 and 92 spots was chosen as the basic printing condition. The concentration of β-Lg probes was 0. 5 mg / mL. The β-Lg detection antibody titre was 1∶2000. One percent no protein blocking solution was choosen as the blocking buffer. The lower detection limit and the biological detection limit of β-Lg were 17. 54 ng / m L and 55. 31 ng / m L respectively. The linear range was determined according to the S type curve of β-Lg and the best fitting models and standard curve were established for β-Lg( R~2=0. 9993).
Conclusion:
The study optimizes conditions of a quantitative analysis system for measurement of β-Lg with protein chip, thus establishing the protein chip platform for quantitative detection of bovine milk β-Lactoglobulin.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP9066459	β-Lactoglobulin B from bovine milk		9066-45-9	Price
IAR4248950	Anti-STUB1/CHIP antibody, mouse monoclonal			Price

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