Homotype cysteine detecting method

Abstract

The present invention relates to a method of producing homocysteine diagnostic kit, which is based on the small molecule capture technology. The method adopts the combination of gene mutation enzyme and small molecule substance to be detected; the gene mutation enzyme is named as small molecule capture enzyme; and the small molecule capture enzyme after mutation and modification maintain and even enhance the affinity to the substrate or the small molecule substance to be detected, but the catalytic capability and the decomposition capacity of the small molecule capture enzyme after mutation and modification to the substrate or the small molecule substance to be detected is weakened. The invention provides a method of researching and producing the homocysteine (HCY) diagnostic kit, which is based on the small molecule capture technology, and also provides mutated S-adenosine homocysteine hydrolase; and the specific S-adenosine homocysteine (SAH) capture enzyme can fully maintain and even enhance the affinity to the S-adenosine homocysteine (SAH), but the catalytic capability to the HCY or the S-adenosyl homocysteine (SAH) is weakened. The method is in particular suitable for small molecule substances, such as inorganic ions, amino acids, polypeptides, nucleotides, oligonucleotide, vitamins, monosaccharides, oligosaccharides, lipids and organic acids.

Description

technical field [0001] The invention relates to components and a detection method, and is mainly aimed at small molecules to be detected. The detection method for homocysteine ​​is developed and produced based on the small molecule capture technology. This method uses a gene mutant enzyme to combine with the small molecule substance to be tested. This gene mutant enzyme is called a small molecule capture enzyme. These mutated and modified small molecules The capture enzyme retains or even enhances its affinity to the substrate or the small molecule substance to be tested, but weakens its ability to catalyze and decompose the affinity to the substrate or the small molecule substance to be tested. The method of developing and producing homocysteine ​​(HCY) diagnostic kit based on small molecule capture technology provides this, and also provides mutant S-adenosyl homocysteine ​​hydrolase, specific S-adenosyl homocysteine Acid (SAH) trapping enzymes can fully retain or even enha...

AI Technical Summary

Problems solved by technology

Polymorphisms in the gene for tetrahydrofolate reductase lead to folic acid deficiency which affects the sensitivity of homocysteine ​​levels (Boers, et al., J. Inher. Metab. Dis., 20:301-306 (1997) )

First, for many methods, specific binders are not easy to prepare and lack specificity, which affects the specificity of detection

Although this defect can be made up by preparing high-molecular-weight antibodies, especially purified monoclonal antibodies, this process takes a lot of time and costs a lot

In addition, for some small-molecular-weight analytes, the option of preparing antibodies becomes infeasible, because small-molecular substances have weak antigenicity

The preparation of small molecular substance antibodies usually requires the combination of large molecular substances and small molecular substances, which leads to more ineffective and more efficient expression of antibodies.

Second, many detection methods, especially for the detection of small molecules, require a lot of time, including chemically induced transformation and chromatographic separation steps

Third, many of these detection methods require the use of sophisticated, expensive specialized analytical instruments such as high-pressure liquid chromatography (HPLC) and gas chromatography (GC / MS)