Kit for detecting Golgi protein 73 by magnetic particle chemiluminescence method and preparation method thereof
A chemiluminescence and Golgi technique, applied in the field of immunoassays, can solve the problems of not requiring tumor screening and not being the basis for early diagnosis or diagnosis of malignant tumors, and achieves simplified experimental operation procedures, increased non-specific interference, high affinity Effect
A chemiluminescence and Golgi technique, applied in the field of immunoassays, can solve the problems of not requiring tumor screening and not being the basis for early diagnosis or diagnosis of malignant tumors, and achieves simplified experimental operation procedures, increased non-specific interference, high affinity Effect
CN113533734APending Publication Date: 2021-10-22TAIZHOU ZECEN BIOTECH CO LTD
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Embodiment 1
[0049] Biotinylated GP73 antibody solution, the preparation steps are as follows:
[0050] 1) Measure 1mg of GP73 antibody (product number ABGP73010 monoclonal antibody, source is mouse antibody);
[0051] 2) Calculate the molecular weight of the antibody (150000) and the molecular weight of biotin-NHS (587), and weigh the biotin-NHS according to the molar ratio of 1:20;
[0052] 3) Weigh 0.4 mg of biotin-NHS and dissolve it with DMSO to a final concentration of 5 mg / ml;
[0053] 4) Add biotin-NHS dissolved in DMSO to the GP73 antibody, mix thoroughly and react at room temperature for 2 hours;
[0054] 5) Dialyzing the reaction product in step 4) with a dialysis bag with a molecular weight cut-off of 500 (Daltons), the dialysis buffer is 0.15M PBS buffer at pH=7.5;
[0055] 6) Dilute the mixture to 0.5 μg / ml with anti-reagent buffer as a biotinylated GP73 antibody solution.
[0056] Alkaline phosphatase-labeled Golgi protein 73 antibody (product number ABGP73020) solution, ...
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The invention discloses a kit for detecting Golgi protein 73 by a magnetic particle chemiluminescence method and a preparation method thereof, and belongs to the technical field of immunoassay. The kit comprises a streptavidin-coated magnetic particle suspension solution, a biotinylation labeled GP73 antibody solution, an alkaline phosphatase labeled Golgi protein 73 antibody solution, a calibrator, a washing solution and a substrate solution. The problems that pathological specimens need to be obtained in histological detection, the GP73 state of a patient who does not obtain cancer tissue pathology cannot be judged, and repeated detection is difficult are solved. Serological detection has the characteristics of convenience, repeatable measurement, quantification, objectivity, real-time detection and the like. The method has the advantages that the reaction time is short, radioactive contamination is avoided, the experimental operation process is simplified, the binding between the biotin and the streptavidin has extremely high affinity, and the reaction has high specificity. Therefore, non-specific interference is not increased while the sensitivity is improved.
Description
technical field [0001] The invention relates to the technical field of immune analysis, in particular to a kit and a preparation method for detecting Golgi protein 73 by magnetic particle chemiluminescence. Background technique [0002] Hepatocellular carcinoma is a malignant tumor with high degree of malignancy, rapid progression, poor prognosis and strong invasiveness. Alpha-fetoprotein (AFP) is an important indicator for the diagnosis of hepatocellular carcinoma, but about 30% of patients with hepatocellular carcinoma are negative for AFP. [0003] Golgi protein 73 (GP73) is a newly discovered hepatocellular carcinoma-related protein. More and more studies have shown that it has important application value in the diagnosis and differential diagnosis of liver cancer and is a potential tumor marker. [0004] Golgi protein 73 (GP73), also known as type II Golgi transmembrane protein (GOLPH2) or Golgi membrane protein I (GOLM1), with a molecular weight of 73KD, is constantly...
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Patent Timeline
22 Oct 2021
Publication
CN113533734A
- IPC
- G01N33/577; G01N33/574; G01N33/68; G01N33/543; G01N33/535; G01N33/532; G01N33/96; G01N21/76
- CPC
- G01N33/577; G01N33/6893; G01N33/57484; G01N33/57438; G01N33/54326; G01N33/535; G01N33/532; G01N33/96
- Inventors
- 许可; 刘振世