Lawson G M, Katzmann J A, Kimlinger T K, O'Brien J F
Clin Chem. 1985 Mar;31(3):381-5.
We have prepared murine monoclonal antibodies against isolated human bone alkaline phosphatase (ALP, EC 3.1.3.1). Hybridoma supernates were separately screened for reactivity against both human liver and bone ALP. Although most antibody-positive hybrids showed similar reactivity against both isoenzymes, one hybridoma produced an antibody that interacted preferentially with liver ALP. This antibody was purified and used to establish an immunoassay to differentiate liver ALP from bone ALP. When equal activities of the two isoenzymes (as determined by a conventional enzymic assay) were measured by the immunoassay, a fivefold greater response was obtained with liver than with bone ALP. The immunoassay can be used to measure the proportions of the bone and liver isoenzymes in mixtures of them. Cross reactivity with human placental and intestinal ALP is less than 3% relative to liver ALP. These findings support the feasibility of developing immunological methods to differentiate these isoenzymes in the clinical laboratory.
我们制备了针对分离出的人骨碱性磷酸酶（ALP，EC 3.1.3.1）的鼠单克隆抗体。分别筛选杂交瘤培养上清液与人肝碱性磷酸酶和骨碱性磷酸酶的反应性。尽管大多数抗体阳性杂交瘤对两种同工酶表现出相似的反应性，但有一个杂交瘤产生了一种优先与人肝碱性磷酸酶相互作用的抗体。该抗体被纯化并用于建立一种免疫测定法，以区分肝碱性磷酸酶和骨碱性磷酸酶。当通过免疫测定法测量两种同工酶的等量活性（通过传统酶法测定）时，肝碱性磷酸酶的反应比骨碱性磷酸酶高五倍。该免疫测定法可用于测量它们混合物中骨和肝同工酶的比例。与人胎盘碱性磷酸酶和肠碱性磷酸酶的交叉反应性相对于肝碱性磷酸酶小于3%。这些发现支持了在临床实验室开发免疫方法区分这些同工酶的可行性。
