CLARiTY光谱仪用于直接测量混浊溶液中的蛋白质浓度

CLARiTY光谱仪用于直接测量混浊溶液中的蛋白质浓度

Label-Free, Direct Measurement of Protein Concentrations in Turbid Solutions with a UV–Visible Integrating Cavity Absorbance Spectrometer

已经广泛研究了蛋白质 – 颗粒缀合物和混合物在生物技术中的多种应用。 然而,缺乏测量蛋白质 – 颗粒溶液的蛋白质浓度的一般方法。 通常,混浊溶液中的蛋白质需要与另一种发色团分离或染色以定量它们的浓度。 在这里,我们展示了使用UV-vis积分腔吸收光谱仪测量混浊溶液中蛋白质浓度的无标记直接方法。 三种系统用于测试测量准确蛋白质浓度的能力:吸附到Alhydrogel的蛋白质,溶液中含有金纳米颗粒的蛋白质,以及包封在聚合物微球体内的蛋白质。 使用由280nm处的吸光度产生的校准曲线成功地量化三种蛋白质 – 颗粒系统中的每一种中的蛋白质浓度。

Protein–particle conjugates and mixtures have been investigated extensively for their diverse applications in biotechnology. However, general methods to measure protein concentration of protein–particle solutions are lacking. Typically, proteins in turbid solutions require separation or staining with another chromophore to quantitate their concentration. Here we demonstrate a label-free, direct approach to measure protein concentrations in turbid solutions using a UV–vis integrating cavity absorbance spectrometer. Three systems are used to test the ability to measure accurate protein concentrations: proteins adsorbed to Alhydrogel, proteins in solution with gold nanoparticles, and proteins encapsulated within polymeric microspheres. Protein concentrations in each of the three protein–particle systems were successfully quantified using a calibration curve created from the absorbance at 280 nm.