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Process and monoclonal antibody for the specific determination of pancreas alpha-amylase in the presence of saliva alpha-amylase

机译:在唾液α-淀粉酶存在下特异性测定胰腺α-淀粉酶的方法和单克隆抗体

摘要

In body fluids containing saliva alpha amylase and pancreatic alpha amylase, pancreatic alpha amylase is determined with a monoclonal antibody which specifically binds but does not inhibit saliva alpha amylase and which has a cross-reactivity of 5% or less toward pancreatic alpha amylase. The monoclonal antibody binds salvia alpha amylase to form a complex which is separated to permit determining pancreatic alpha amylase with an amylase detection system. The complex may be separated by precipitating with a precipitating agent such as an anti-antibody or protein A, or by immobilizing the monoclonal antibody on a solid carrier. The monoclonal antibody is preferably produced by immunizing a host animal with an immunogen containing aluminum hydroxide, Bordatella pertussis and native saliva alpha amylase at least 7 times over a period of at least 9 months, fusing B lymphocytes of the immunized host animal with a transforming agent to produce hybridomas, cloning and culturing the hybridomas, and screening antibodies formed.
机译:在含有唾液α淀粉酶和胰α淀粉酶的体液中,用特异性结合但不抑制唾液α淀粉酶且对胰腺α淀粉酶的交叉反应率为5%以下的单克隆抗体测定胰α淀粉酶。单克隆抗体与丹参α淀粉酶结合形成复合物,将其分离以允许使用淀粉酶检测系统测定胰腺α淀粉酶。可以通过用沉淀剂如抗抗体或蛋白A沉淀或通过将单克隆抗体固定在固体载体上来分离复合物。优选通过在至少9个月的时间内用含有氢氧化铝,百日咳博德特氏菌和天然唾液α淀粉酶的免疫原免疫宿主动物至少7次,并用转化剂融合免疫的宿主动物的B淋巴细胞来产生单克隆抗体。产生杂交瘤,克隆和培养杂交瘤,并筛选形成的抗体。

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