Division of Rheumatology and Arthritis Centre, Department of Medicine, Queen's University, Kingston, ON K7L3N6, Canada.
Anal Biochem. 2010 Jan 15;396(2):310-2. doi: 10.1016/j.ab.2009.09.037. Epub 2009 Sep 24.
The quantitation of low to moderately abundant serum proteins is a common problem encountered in biochemistry. A physical property of the antigen of interest needs to be exploited in the initial binding step of an immunoassay resulting in capture (purification). We describe a two-stage immunoassay utilizing chromogenic and chemiluminescent substrates which is applied to two serum anionic proteins. In this assay, anionic serum proteins are selectively bound to positively charged chitosan-coated polystyrene plates at pH 6.1, in the presence of detergent. The assay has a detection limit of 0.1 microg/mL and a 1000-fold range.
定量检测低丰度和中丰度血清蛋白是生物化学中常见的问题。免疫测定的初始结合步骤需要利用感兴趣抗原的物理性质，从而实现捕获（纯化）。我们描述了一种两步免疫测定法，该方法利用显色和化学发光底物，应用于两种血清阴离子蛋白。在该测定中，在去污剂存在的情况下，阴离子血清蛋白在 pH 值为 6.1 时被选择性地结合到带正电荷的壳聚糖涂覆的聚苯乙烯板上。该测定法的检测限为 0.1 μg/mL，检测范围为 1000 倍。
