Buffone Gregory J
Department of Clinical Laboratories, Children's Hospital National Medical Center and George Washington University, Washington, D.C. 20009.
J Res Natl Bur Stand A Phys Chem. 1976 Jul-Aug;80A(4):605-608. doi: 10.6028/jres.080A.059. Epub 1976 Aug 1.
Light scattering methods for the physical analysis of synthetic and biological polymers necessitates the use of scattering standards and absolute light scattering measurements. Standardization has not been employed when light scattering has been used to monitor immunochemical reactions using a kinetic or thermodynamic mode. The concentration of a specific protein present in a complex matrix such as urine, serum or cerebrospinal fluid, is measured by reacting the protein of interest with its specific antibody and then measuring the excess light scattering of the solution produced by the formation of antigen antibody complexes. The lack of established light scattering standards in the area of immunochemical measurements makes instrumental quality control difficult and has hindered direct comparison of data among investigators. Both solid and liquid light scattering standards would be necessary to encompass the wide range of instrumentation currently in use. Several solid standards which have been used in the past include reflecting diffusers such as vitrolite, magnesium carbonate crystals with a ground surface, magnesium oxide coatings on magnesium carbonate crystal, casein paint on vitrolite, and solid opal glass transmitting diffusers such as flashed opal glass and solid opal glass. These standards, while applicable to manual light scattering photometers, are not suitable for recently developed automated instrumentation. Liquid standards in the form of Ludox, solutions of polystyrene, suspensions of small diameter latex spheres and even pure organic solvents could be used more easily with the continuous flow and discrete automated analyzers. The introduction of instrumental standards at this level of analysis would result in improved overall quality control and facilitate data and method comparison between laboratories.
用于合成聚合物和生物聚合物物理分析的光散射方法需要使用散射标准品和绝对光散射测量。当使用光散射以动力学或热力学模式监测免疫化学反应时,尚未采用标准化方法。通过使目标蛋白质与其特异性抗体反应,然后测量由抗原抗体复合物形成产生的溶液的过量光散射,来测量存在于复杂基质(如尿液、血清或脑脊液)中的特定蛋白质的浓度。免疫化学测量领域缺乏既定的光散射标准品,使得仪器质量控制变得困难,并阻碍了研究人员之间数据的直接比较。固体和液体光散射标准品对于涵盖目前使用的广泛仪器设备都是必要的。过去使用过的几种固体标准品包括反射漫射器,如玻璃彩石、表面研磨的碳酸镁晶体、碳酸镁晶体上的氧化镁涂层、玻璃彩石上的酪蛋白漆,以及固体蛋白石玻璃透射漫射器,如闪光蛋白石玻璃和固体蛋白石玻璃。这些标准品虽然适用于手动光散射光度计,但不适用于最近开发的自动化仪器。以硅石溶胶、聚苯乙烯溶液、小直径乳胶球悬浮液甚至纯有机溶剂形式存在的液体标准品,在连续流动和离散自动分析仪中使用起来会更容易。在这个分析层面引入仪器标准品将提高整体质量控制,并促进实验室之间的数据和方法比较。