公开(公告)号：US5187107A

公开(公告)日：1993-02-16

申请号：US722034

申请日：1991-06-27

申请人： Michael I. Watkins ,  Clifford R. Bartlett ,  Edward T. Liang ,  John M. Pocekay ,  Mark A. Staples

发明人： Michael I. Watkins ,  Clifford R. Bartlett ,  Edward T. Liang ,  John M. Pocekay ,  Mark A. Staples

IPC分类号： G01N33/53 ,  G01N33/531 ,  G01N33/82

CPC分类号： G01N33/82 ,  Y10T436/25 ,  Y10T436/25125

摘要： Denaturing agents used to pretreat test samples for immunoassays are neutralized and deactivated by the addition of a treatment agent which is a combination of a buffering agent in acid form and an agent for converting the sulfhydryl groups of the denaturing agents to a non-active form such as disulfides or alkylthio groups. The treatment agent serves this function without having sufficient denaturing activity by itself to interfere with subsequent steps of the assay procedure. Also disclosed is a protocol for a vitamin B.sub.12 assay which involves a sequential rather than competitive binding of the B.sub.12 with excess intrinsic factor followed by the binding of excess enzyme-labeled B.sub.12 to any remaining intrinsic and immobilizing such intrinsic factor on a solid phase. Various problems associated with B.sub.12 assays are avoided by this technique.

摘要翻译： 用于预处理试验样品用于免疫测定的变性剂通过加入处理剂中和和失活，所述处理剂是酸形式的缓冲剂和用于将变性剂的巯基转化成非活性形式的试剂 作为二硫化物或烷硫基。 治疗剂不具有足够的变性活性来干扰随后的测定程序步骤而起作用。 还公开了一种用于维生素B12测定的方案，其涉及B12与过量内在因子的顺序而非竞争性结合，然后将过量的酶标记的B12与任何残留的内在和固定在固相上的固有因子结合。 通过这种技术避免了与B12测定相关的各种问题。