申请人:Abbott Laboratories
公开号:US05843634A1
公开(公告)日:1998-12-01
This invention relates to genetically engineered enzymes, their ligand conjugates, their manufacture, and their use in qualitative or quantitative assays. A hybrid enzyme, such as an AP-epitope, has a foreign amino acid moiety (an epitope) inserted near the active site of the starting AP enzyme. The foreign amino acid moiety binds with an analyte, and, as a consequence of this binding, the enzymatic activity of the hybrid enzyme, AP-epitope, is modified. The changes in the enzymatic activity are dependent upon the presence, or the amount, of the analyte. In another embodiment, the hybrid enzyme consists of a cysteine introduced near the active site of an AP to give a hybrid enzyme. The cysteine on the hybrid enzyme serves as a point of conjugation of a ligand, such as theophylline, ferritin, thyroxine, or digoxigenin, to form the hybrid enzyme-ligand conjugate. The ligand binds with an antibody, an analyte or a binding molecule to an analyte and as a result of this binding, the enzymatic activity of the hybrid enzyme-ligand conjugate is modified or modulated.
这项发明涉及基因工程酶、它们的配体缀合物、它们的制造方法以及它们在定性或定量分析中的应用。一种杂交酶,如AP-表位酶,在起始AP酶的活性位点附近插入了一个外源氨基酸残基(一个表位)。外源氨基酸残基与一个分析物结合,由于这种结合,杂交酶AP-表位的酶活性被改变。酶活性的变化取决于分析物的存在或数量。在另一实施例中,杂交酶由在AP的活性位点附近引入的一个半胱氨酸形成。杂交酶上的半胱氨酸作为配体(如茶碱、铁蛋白、甲状腺素或地高辛)缀合的位点,以形成杂交酶-配体缀合物。配体与抗体、分析物或与分析物结合的分子结合,由于这种结合,杂交酶-配体缀合物的酶活性被改变或调节。