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Preparation method of quality control serum for ischemia-modified albumin
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A protein and serum technology, applied in the field of preparation of ischemia-modified albumin control products, can solve problems such as separation and purification difficulties, and achieve the effects of simple preparation process, avoiding matrix effect, and sufficient sources
Active Publication Date: 2013-09-18
NINGBO MEDICAL SYSTEM BIOTECHNOLOGY CO LTD
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Problems solved by technology
Due to the particularity of the nitrogen-terminal sequence of HSA, the possibility of obtaining human IMA from other animals is ruled out, and because the difference between IMA and HSA is very small, there may be only 2-4 amino acid differences, so it is also very difficult to separate and purify it from HSA. difficulty
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Embodiment 1
[0025] (1) Collect 100 ml of serum from healthy people who have excluded infectious diseases and common diseases, and use it as a serum raw material to adjust the pH value to 7.2;
[0026] (2) Add 9.2 mM FeSO 4 and 39 mM hydrogen peroxide;
[0027] (3) Reaction under UV light irradiation at 37°C for 20 min;
[0028] (4) Dialyze in 10 mM Tris-HCl buffer at 25°C, pH 7.5, for 12 h;
[0029] (5) Add 0.1 ml of 4-chloro-3,5-dimethylphenol, mix evenly, and divide into glass bottles. The volume of each bottle is 1 ml, and the ELISA kit is used to determine its value.
Embodiment 2
[0031] (1) Collect 100 ml of serum from healthy people who have excluded infectious diseases and common diseases, and use it as a serum raw material to adjust the pH value to 7.5;
[0032] (2) Add 1 mM iron porphyrin and 10 mM hydrogen peroxide;
[0033] (3) React at 37°C for 30 minutes under visible light mercury lamp irradiation;
[0034] (4) Dialyze in 25°C, pH 8.0, 100 mM disodium hydrogen phosphate-citrate buffer for 24 h;
[0035] (5) Add 0.1 g NaN 3 , mixed evenly, and distributed in glass bottles, the volume of each bottle was 1 ml, and the ELISA kit was used to determine its value.
Embodiment 3
[0037] (1) Collect 100 ml of serum from healthy people who have excluded infectious diseases and common diseases, and use it as a serum raw material to adjust the pH value to 7.0;
[0038] (2) Add 0.5 mM CuSO 4 and 50 mM hydrogen peroxide;
[0039] (3) React for 15 minutes under visible light mercury lamp irradiation at 37°C;
[0040] (4) Dialyze in 100 mM imidazole buffer at 25°C, pH 7.8, for 18 h;
[0041] (5) Add 0.1 ml of 4-chloro-3,5-dimethylphenol, mix evenly, and divide into glass bottles. The volume of each bottle is 1 ml, and the ELISA kit is used to determine its value.
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Abstract
The invention discloses a preparation method of quality control serum for ischemia-modified albumin; the preparation method comprises the following steps of: (1) adopting the blood serum of a healthy person, and adjusting a pH value to be 7.0-7.5; (2) adding 0.1-100mm of metal ionic salt with a variable valence center or a metal organic composite with molecule planarity to the blood serum, and then adding 0.1-100mm of hydrogen peroxide; (3) reacting for 10-30min under the 35-40 DEG C illumination; (4) dialyzing a solution obtained through the reaction in the step (3) in 0.1-1000mm of a dialysis buffer solution with pH of 7.5-8.0 under the 20-25 DEG C temperature for 12-24h; and (5) adding a preservative according to a conventional technology, subpackaging and determining after mixing uniformly. The preparation method has the advantages of adequate the source of raw materials, easily obtaining of the raw materials, simple preparation technology, few amount of additives, low production cost, small non-specificity interference, no needing for composite dissolving, good homogeneity and small difference between bottles.
Description
technical field [0001] The invention relates to a preparation method of an ischemic modified albumin control product. Background technique [0002] Acute coronary syndrome (ACS) has different clinical manifestations, but they all have the same pathophysiological process, that is, atherosclerotic plaque shedding, platelet aggregation and thrombus formation, leading to coronary artery stenosis and blockage. And cause myocardial ischemia, if the myocardial ischemia time is prolonged, it will lead to myocardial cell necrosis and myocardial injury. Therefore, identification of myocardial ischemia before myocardial necrosis, early diagnosis and initiation of treatment are crucial for patients. At present, the diagnosis of cardiac ischemia mostly relies on the changes of the ST segment and T wave of the electrocardiogram, which is the easiest, fastest and cheapest examination method for myocardial ischemia, but patients with myocardial infarction may also show no abnormalities in ...
Claims
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