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    • 4. 发明申请
    • GENETICALLY ENGINEERED ENZYMES AND THEIR CONJUGATES FOR DIAGNOSTIC ASSAYS
    • 基因工程酶及其与诊断学结合的诊断
    • WO1994020636A1
    • 1994-09-15
    • PCT/US1994002539
    • 1994-03-09
    • ABBOTT LABORATORIES
    • ABBOTT LABORATORIESBRATE, Elaine, M.BRENNAN, Catherine, A.BRIDON, Dominique, P.JAFFE, Keeve, D.KRAFFT, Grant, A.MANDECKI, WlodzimierzMARCH, Steven, C.RUSSELL, John, C.YUE, Vincent, T.
    • C12Q01/00
    • C07K14/59C07K2319/40C12N9/16C12Q1/42
    • 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.
    • 本发明涉及基因工程改造的酶,其配体缀合物,其制备及其在定性或定量测定中的用途。 诸如AP-表位的杂交酶在起始AP酶的活性位点附近插入外源氨基酸部分(表位)。 外来氨基酸部分与分析物结合,并且作为该结合的结果,修饰了杂合酶AP-表位的酶活性。 酶活性的变化取决于分析物的存在或数量。 在另一个实施方案中,杂交酶由在AP的活性位点附近引入的半胱氨酸组成以产生杂交酶。 杂交酶上的半胱氨酸用作配体如茶碱,铁蛋白,甲状腺素或地高辛的缀合点,以形成杂交酶 - 配体缀合物。 配体与分析物结合抗体,分析物或结合分子,作为该结合的结果,修饰或调节杂交酶 - 配体结合物的酶活性。