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Preparation method of antithrombin III activity assay kit

An activity assay and antithrombin technology, which is applied in the field of preparation of antithrombin III activity assay kits, can solve the problems of high cost, prone to errors in results, inconvenient operation and the like, and achieves low cost and good stability. , the effect of simple operation

Inactive Publication Date: 2016-06-01
CHENGDU XIEHE BIOLOGICAL TECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

This technology is expensive, and the operation is inconvenient, and the result is prone to error

Method used

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  • Preparation method of antithrombin III activity assay kit
  • Preparation method of antithrombin III activity assay kit

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Embodiment Construction

[0019] In conjunction with embodiment, further set forth the present invention:

[0020] The embodiment takes the preparation of the ATIII activity detection reagent and the specific detection on the automatic coagulation apparatus CA-1500 as an example.

[0021] (1) Use 10-50mmol / LTris-HCl, pH8.4 solution to dissolve thrombin freeze-dried powder and heparin sodium solution, mix different concentrations of thrombin and heparin sodium solution, and prepare S1 reagent, the thrombin concentration is 5 ~20IU / mL, the heparin concentration is 0.2~10IU / mL.

[0022] (2) Dissolve the chromogenic substrate S-2238 or Sar-Pro-Arg-P-nitroanilide with 10-50 mmol / LTris-HCl of pH 7.0, and prepare a solution with a substrate concentration of 2-5 μmol / mL, as S2 reagent.

[0023] (3) Add stabilizers and excipients to S1 reagent and S2 reagent respectively and freeze-dry to prepare S1 freeze-dried product and S2 freeze-dried product. The content of additives added to S1 reagent in S1 reagent i...

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Abstract

The invention relates to a preparation method of a clinical detection reagent and in particular relates to a preparation method of an antithrombase III activity determination kit. The preparation method of the antithrombase III activity determination kit comprises the following steps: dissolving lyophilized antithrombase powder and sodium heparin with Tris-HCl to form an S1 reagent, dissolving antithrombase chromogenic substrate S-2238 or Sar-Pro-Arg-P-nitroanilide with Tris-HCl to form an S2 reagent, lyophilizing the reagents S1 and S2 to form an S1 lyophilized product and an S2 lyophilized product; redissolving the S1 lyophilized product with Tris-HCl containing sodium azide solution to form an S1 lyophilized and redissolved reagent; dissolving the S2 lyophilized product with ultrapure water to form an S2 lyophilized and redissolved reagent; lyophilizing multi-person mixed plasma to prepare an ATIII calibrator; valuing the ATIII calibrator in the range of 90-110%; drawing a standard curve by adopting the valued ATIII calibrator, the S1 hyophilized and redissolved reagent and the S2 hyophilized and redissolved reagent, wherein linear correlation coefficient is larger than 0.99; and adding the S1 hyophilized and redissolved reagent and the S2 hyophilized and redissolved reagent into a to-be-detected plasma sample, and carrying out chromogenic reaction. The antithrombase III activity determination kit obtained by adopting the preparation method provided by the invention is low in cost, wide in linear range, good in stability and easy to use.

Description

technical field [0001] The invention relates to a preparation method of a clinical detection reagent, in particular to a preparation method of an antithrombin III activity assay kit. Background technique [0002] Antithrombin III (antithrombin III, ATIII) is an inhibitor of thrombin and factors XIIα, XIα, IXα, Xα and other serine-containing proteases. It binds to thrombin via an arginine-serine peptide bond. The ATIII thrombin complex is formed to inactivate the enzyme, and heparin can accelerate this reaction by more than a thousand times. The combination of heparin and the lysine contained in ATIII causes the conformation change of ATIII, which makes the arginine residue contained in ATIII easier to bind with the serine residue of thrombin. Once the heparin-ATIII thrombin complex is formed, heparin dissociates from the complex and recombines with another molecule of ATIII for repeated utilization. The ATIII-thrombin complex is eliminated by the reticuloendothelial syste...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/56
CPCG01N33/86G01N2333/8128
Inventor 肖薇林方昭谢兆林杨显福孙小成肖玲
Owner CHENGDU XIEHE BIOLOGICAL TECH
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