申请人:TOSOH CORPORATION
公开号:EP0347256A2
公开(公告)日:1989-12-20
A method for detecting a substance in a sample which comprises steps of:
(a) mixing a first enzyme complex comprising a first enzyme bound to a material having the affinity for the substance to be detected with the sample,
(b) separating the first enzyme complex bound to the substance to be detected (the first enzyme complex in bound form) from the first enzyme complex not bound to the substance to be detected (the first enzyme complex in free form),
(c) reacting the first enzyme complex in bound form with (i) a first substrate comprising a substrate for said first enzyme which is bound to a second enzyme; or (ii) a first substrate comprising a substrate to said first enzyme which is not bound to a second enzyme,
(d) removing the unreacted first substrate from the first product formed in step (i); or
reacting the first product formed in step (ii) with a second enzyme complex comprising a second enzyme bound to a material having the affinity for said first product, and then removing the unreacted second enzyme complex,
(e) reacting the system with a second substrate to the second enzyme to form a second product, and
(f) detecting the second product produced in step (e).
一种检测样品中物质的方法,包括以下步骤
(a) 将第一酶复合物与样品混合,第一酶复合物包括与待检测物质具有亲和力的 材料结合的第一酶、
(b) 将与待检测物质结合的第一种酶复合物(结合形式的第一种酶复合物)与未与待检测物质结合的第一种酶复合物(游离形式的第一种酶复合物)分离、
(c) 使结合形式的第一酶复合物与 (i) 第一底物反应,该底物包括与第二酶结合的所述第一酶的底物;或 (ii) 第一底物反应,该底物包括与第二酶未结合的所述第一酶的底物、
(d) 从步骤(i)中形成的第一产物中除去未反应的第一底物;或
将步骤(ii)中形成的第一产物与第二酶复合物反应,第二酶复合物包括与对所述第一产物具有亲和力的材料结合的第二酶,然后除去未反应的第二酶复合物、
(e) 用第二种底物与第二种酶反应,形成第二种产物,以及
(f) 检测步骤(e)中产生的第二产物。