[Optimizing the detection of bovine milk β-Lactoglobulin with protein chip].

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.