Ithakissios D S, Kubiatowicz D O
Clin Chem. 1977 Nov;23(11):2072-9.
We describe a radioassay method that involves the use of magnetic protein microparticles composed of a water-insoluble protein matrix containing magnetically responsive material. We define two different types of particles according to the mechanism of action: (a) The substrate is sorbed nonspecifically by the protein matrix of the particle or by a second substance such as charcoal or ion-exchange resin incorporated within the protein matrix of the particle. These particles are useful for separating free from bound substrate. Examples of these are albumin magnetic microparticles for use in a total thyroxine radioassay and triiodothyronine uptake test, or albumin magnetic microparticles containing charcoal for use in a vitamin B12 radioassay. (b) The substrate is sorbed specifically by a binding protein incorporated within the matrix of the particles. The binding protein can include antibodies or other specific nonimmune proteins. Particles of this type are useful in solid-phase radioassays. These particles are exemplified by albumin magnetic microparticles containing sockeye salmon serum, used in a solid-phase B12 radioassay. We discuss the methods for the preparation of both types of magnetic microparticles and their use in radioassays. We describe a unique inexpensive magnetic separation rack, which provides simple, fast, and reproducible separation of the magnetic microparticles from their suspending medium during the assay.
我们描述了一种放射分析方法,该方法涉及使用由含有磁响应材料的水不溶性蛋白质基质组成的磁性蛋白质微粒。我们根据作用机制定义了两种不同类型的微粒:(a) 底物通过微粒的蛋白质基质或通过掺入微粒蛋白质基质中的第二种物质(如木炭或离子交换树脂)被非特异性吸附。这些微粒可用于分离游离底物和结合底物。此类例子包括用于总甲状腺素放射分析和三碘甲状腺原氨酸摄取试验的白蛋白磁性微粒,或用于维生素B12放射分析的含木炭的白蛋白磁性微粒。(b) 底物通过掺入微粒基质中的结合蛋白被特异性吸附。结合蛋白可包括抗体或其他特异性非免疫蛋白。此类微粒可用于固相放射分析。此类微粒的例子是用于固相B12放射分析的含红大马哈鱼血清的白蛋白磁性微粒。我们讨论了这两种磁性微粒的制备方法及其在放射分析中的应用。我们描述了一种独特的廉价磁性分离架,它在分析过程中能将磁性微粒与其悬浮介质简单、快速且可重复地分离。