很多研究工作都需要对蛋白质进行纯化，只有经过纯化后的蛋白质方可用于后续的生物物理学，结构学，诊断治疗等方面的研究。然而，很多研究者常常忽视蛋白质纯化后的质量控制环节，结果导致实验数据重复性差或者得到的结论是错误的。因此蛋白质纯化后在用于后续实验前必须经过检测与定量。这篇文章主要简单介绍蛋白质纯化后分析过程中所用到的一系列技术。
检测纯度与完整度
- 电泳：聚丙烯酰胺凝胶电泳（SDS-PAGE）是一种快速简单的监测蛋白质样品纯度的技术手段，最少能够监测到100 ng的蛋白质。除了SDS-PAGE外，等电聚焦电泳（IEF）以及毛细管电泳（CE）也能够用来分离蛋白质。
- 紫外可见光谱：能够用来监测纯化蛋白质中的主要污染物核酸。核酸在260nm处有吸收值。对于没有核酸污染的蛋白质样品，260/280 nm吸光度的比值应该接近0.57。
- 质谱：质谱主要用来监测蛋白质的完整度。质谱能够利用很少量的样品准确的测量蛋白质的分子量，其最大测量值可达到500 KD。另外，质谱也能够提供蛋白质翻译后修饰的信息，比如磷酸化，乙酰化，糖基化，泛素化等。
检测均一性
- 动态光散射（DLS）：DLS是一种用来监测蛋白质是否形成聚集体的手段。能够监测蛋白质是否虽时间及温度的变化发生聚集。
- 紫外可见光与荧光光谱: 紫外可见光光谱可以用来检测蛋白质的均一性。直径大于200nm的颗粒在大于320nm波长范围会产生吸光值。激发光波长在280 nm, 测量样品在280 nm与340 nm发射光强度的比值，这一比值与蛋白质的聚集情况具有直接相关性。比值为0时，样品中几乎不含任何聚集体，比值为１时，样品中有明显聚集发生。
- 尺寸排阻色谱(SEC): SEC是目前定量蛋白质多聚体的标准分离技术。在尺寸排阻色谱中更大体积的组分将比小体积组分更先被洗脱。
检测活性
- 总蛋白质浓度: Bradford, BCA, Lowry方法利用标准曲线定量蛋白质浓度。紫外可见光光谱能够用来测量蛋白质浓度，前提是蛋白质的消光系数（extinction coefficient）已知。280nm处的消光系数可以根据蛋白质内氨基酸的组成进行计算得到。蛋白质浓度检测方法见历史文章
- 具有活性的蛋白质浓度: 纯化出的蛋白质样品中活性组分的比重有时非常低。对于结合类蛋白质，表面等离子共振(SPR)是一种很方便的测定活性组分浓度的技术。对于其他类蛋白质，需要根据其活性检测手段选择相应技术。
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