Preparation method of lyophilization type determination PT reagent

A technology of freeze-dried protective agent and reagent, which is applied in the field of preparation of freeze-dried PT reagents, can solve the problems of insufficient protection effect and obvious prolongation of validity period, and achieve easy production, overcome the problem of loss of thromboplastin, and high The effect of clinical application value

Inactive Publication Date: 2019-10-18
山东艾科达生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Most of the current freeze-dried PT reagents only use a freeze-drying protective agent that can protect the freeze-drying effect,

Method used

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  • Preparation method of lyophilization type determination PT reagent
  • Preparation method of lyophilization type determination PT reagent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Including the following steps:

[0021] Q1. Add 3% sodium glutamate, 10% inulin oligosaccharides, and 1% bovine serum albumin into the prepared HEPES buffer. The pH of the HEPES buffer is between 6.4-7.4 In between, stir for 15 minutes to obtain a uniform protective agent solution;

[0022] Q2, dissolving an appropriate amount of synthetic phospholipids in the HEPSE buffer solution containing 1% triton, after stirring for 120 minutes, to obtain a uniformly dispersed phospholipid solution, the mass fraction of the surfactant is 1.0%;

[0023] Q3. Express sheep tissue factor in Escherichia coli by genetic engineering, prepare sheep tissue factor with a purity of more than 90%, and dissolve the sheep tissue factor in an appropriate amount of PBS buffer to form a 55ug / ml recombinant sheep tissue factor solution;

[0024] Q4. Add recombinant sheep tissue factor solution and phospholipid solution, the mass ratio of phospholipid to recombinant sheep tissue factor is 2.8×10 3...

Embodiment 2

[0027] Including the following steps:

[0028] Q1. Add 5% sodium glutamate, 5% inulin oligosaccharides, and 2% bovine serum albumin into the prepared HEPES buffer. The pH of the HEPES buffer is between 6.4-7.4 In between, stir for 15 minutes to obtain a uniform protective agent solution;

[0029] Q2, dissolving an appropriate amount of synthetic phospholipids in the HEPSE buffer solution containing 1% triton, after stirring for 120 minutes, to obtain a uniformly dispersed phospholipid solution, the mass fraction of the surfactant is 1.5%;

[0030] Q3. Express sheep tissue factor in Escherichia coli by genetic engineering, prepare sheep tissue factor with a purity of more than 90%, and dissolve the sheep tissue factor in an appropriate amount of PBS buffer to form a 55ug / ml recombinant sheep tissue factor solution;

[0031] Q4. Add recombinant sheep tissue factor solution and phospholipid solution, the mass ratio of phospholipid to recombinant sheep tissue factor is 2.8×10 3 ...

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Abstract

The invention provides a preparation method of a lyophilization type determination PT reagent. Raw materials comprise HEPES buffer solution, synthetic phospholipids, a recombinant sheep tissue factor,a lyoprotectant and a preservative. The preparation method comprises the following steps: adding the lyoprotectant into prepared HEPES buffer solution, dissolving an appropriate amount of synthetic phospholipids in HEPES buffer solution containing 1% triton X-100, dissolving a sheep tissue factor in an appropriate amount of PBS buffer solution to form 55[mu]g/ml tissue factor solution, adding therecombinant sheep tissue factor into phospholipid solution, finally adding the preservative and lyoprotectant solution into phospholipid tissue factor solution, and performing lyophilization on the obtained supernate by virtue of the conventional technology, thus the PT reagent product is obtained. The preparation method provided by the invention optimizes a formula of a prepared thromboplastin lyoprotectant on the basis of existing components, the problem of loss of thromboplastin in an existing prothrombin detection reagent lyophilizing process is solved, sensitivity is relatively high, a validity period of the reagent is prolonged, and production is easy. The reagent has high clinical application value and potential market benefit.

Description

technical field [0001] The invention belongs to the technical field of biomedical detection, and in particular relates to a preparation method of a freeze-dried PT reagent for determination. Background technique [0002] Tissue factor (TF) is a transmembrane glycoprotein synthesized by adventitial fibroblasts. After vascular injury, when blood contacts subendothelial cells, factor VII combines with TF in the presence of calcium ions to form a bimolecular complex. Causes blood coagulation, TF is a single-chain protein containing 263 amino acids, a signal peptide with a synthetic size of 32 amino acids, and the extracellular or cell surface domain of TF is 219 residues, including three Trp-Lys-Ser repeats sequence, it also contains two disulfide bridges linking Cys 49 to Cys57 and Cys 186 to Cys 209. [0003] Rabbit brain thromboplastin has become a hot spot in the development of (PT) diagnostic reagents in recent years due to its low side effects, clear activity, and strong ...

Claims

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Application Information

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IPC IPC(8): G01N33/535G01N1/28G01N1/42
CPCG01N33/535G01N1/28G01N1/42
Inventor 谯兴强韩墨王醒谭柏清赵志敏
Owner 山东艾科达生物科技有限公司
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