Schwenzer B, Neef B, Bublitz R, Ehle H, Horn A
Biomed Biochim Acta. 1987;46(1):15-21.
Electrophoretically and immunologically homogeneous alkaline phosphatase (AP) can be obtained by a simple and rapid two-step procedure. Step 1: Immunosorbent affinity chromatography using immobilized polyclonal anti-AP-antibodies and elution of the bound enzyme by triethylamine solution, pH 11.4. After affinity chromatography the specific activity of the extracted crude material increases from 3.5 to 793 IU/mg. Step 2: Final purification to a specific activity of 1524 IU/mg by DEAE-cellulose ion exchange chromatography. One investigator is able to purify 100 mg AP within 3 days. The overall recovery is 65%. Methods for characterization and selection of anti-AP-antiserum using immunoinhibition of AP are described.
通过一个简单快速的两步法可获得电泳和免疫均一的碱性磷酸酶（AP）。步骤1：使用固定化多克隆抗AP抗体进行免疫吸附亲和层析，并用pH 11.4的三乙胺溶液洗脱结合的酶。亲和层析后，提取的粗品的比活性从3.5 IU/mg增加到793 IU/mg。步骤2：通过DEAE-纤维素离子交换层析最终纯化至比活性为1524 IU/mg。一名研究人员能够在3天内纯化100 mg AP。总回收率为65%。描述了使用AP的免疫抑制来表征和选择抗AP抗血清的方法。
