Method for detecting ADAAMTS13 enzyme activity in serum by SELDI-TOF-MS (surface-enhanced laser desorption/ionization-time of flight-mass spectrometry)

Abstract

The invention discloses a method for detecting the ADAAMTS13 enzyme activity in serum by SELDI-TOF-MS (surface-enhanced laser desorption / ionization-time of flight-mass spectrometry). The method comprises the following steps of (1) performing ADAAMTS13 enzyme digestion substrate GST-vWF73-6XHis recombination protein expression and purification; (2) taking an enzyme digestion reaction; (3) after an ADAAMTS13 enzyme digestion product is enriched by an IMAC chip, performing recognition and detection on the ADAAMTS13 enzyme digestion product; obtaining the enzyme digestion activity of ADAAMTS13 in the serum sample through analysis software. The detection method is fast and effective; the detection is precise; after the detection on all TMA (thrombotic microangiopathy) suspected cases, all cases with the ADAAMTS13 enzyme activity being less than 10 percent are screened out; the patients are treated as TTP (thrombotic thrombocytopenic purpura) patients. For all TTP cases, in the treatment process and the long-time monitoring after the hospital leaving, the regular detection of the ADAAMTS13 enzyme activity needs to be performed, so that the patient can be better and correctly treated; the treatment cost is reduced; the survival rate and the life quality of the patient are effectively improved.

Description

technical field [0001] The invention belongs to the technical field of medical molecular biology and protein polypeptide detection, and particularly relates to a method capable of detecting metalloprotease ADAMTS13 enzyme activity by using SELDI-TOF-MS, which is applied to TTP disease screening in TMA disease and TTP disease Treatment monitoring. Background technique [0002] SELDI-TOF-MS (Surface Enhanced Laser Desorption Ionization Time-of-Flight Mass Spectrometry) is a new type of mass spectrometry instrument developed by Ciphergen Corporation of the United States. It mainly consists of three parts: the first part is the protein chip (Protein Chip), which is its core technology; The second part is the time-of-flight mass spectrometer (TOF MS), which can also be called the chip reader (protein chip reader); the third part is the system-specific analysis software. [0003] The protein chip is the core technology of SELDI-TOF-MS. This detection method will use the metal aff...

AI Technical Summary

Problems solved by technology

The clinical manifestations of these two diseases are very similar, but one of the biggest differences between the two is the significant difference in the enzymatic activity of metalloproteinase (ADAMTS13)

Images