Genetically engineered enzymes and their conjugates for...

C - Chemistry – Metallurgy – 12 – N

Patent

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C12N 15/52 (2006.01) C07K 14/59 (2006.01) C12N 9/00 (2006.01) C12N 9/16 (2006.01) C12N 15/55 (2006.01) C12Q 1/00 (2006.01) C12Q 1/42 (2006.01) G01N 33/53 (2006.01) G01N 33/531 (2006.01) G01N 33/58 (2006.01)

Patent

CA 2157767

This invention relates to genetically enzymes, their ligand conjugates, their manufactures, 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.

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