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Method for stabilizing activity of alpha-hydroxybutyricdehydrogenaseand lactic dehydrogenase of quality-control serum

A hydroxybutyrate dehydrogenase and lactate dehydrogenase technology, applied in biological testing, material testing and other directions, can solve the problems of high price, short half-life of enzyme molecules, low serum enzyme activity, etc. Inexpensive effect

Active Publication Date: 2010-11-24
AILEX TECH GRP CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] However, at present, the fixed-value quality control serum used in domestic clinical chemistry uses metal ions and sulfhydryl compounds with short stability periods as enzyme stabilizers, resulting in low serum enzyme activity and short half-life of enzyme molecules. There are only a handful of units of fixed-value freeze-dried quality control serum, and the freeze-dried quality control serum produced by foreign companies can be used in clinical chemistry test items, but the price is expensive

Method used

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  • Method for stabilizing activity of alpha-hydroxybutyricdehydrogenaseand lactic dehydrogenase of quality-control serum
  • Method for stabilizing activity of alpha-hydroxybutyricdehydrogenaseand lactic dehydrogenase of quality-control serum
  • Method for stabilizing activity of alpha-hydroxybutyricdehydrogenaseand lactic dehydrogenase of quality-control serum

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] To the phosphate buffer (pH 7.5, concentration 50mmol / L) containing 3% BSA, 200mM glycine, α-HBDH and LDH concentrations of 400U / L and 550U / L respectively, add according to the following formula:

[0034] Among them: the concentration of phosphate buffer saline, by weighing disodium hydrogen phosphate (Na 2 HPO 4 12H 2 O) 6.02g and sodium dihydrogen phosphate (NaH 2 PO 4 2H 2 (0) 0.5g is a benchmark, and the concentration after dissolving and constant volume in distilled water to 1000mL is 50mmol / L;

[0035] (1) No added sugar and sugar alcohol protective agent; (2) 10% mannitol; (3) 10% trehalose; (4) 10% sucrose.

[0036] Take 2 mL of the prepared solution of each group and put it into a brown vial for freeze-drying. Freeze-drying conditions: after vacuuming at a freezing temperature of -50°C, carry out main drying at -20°C for 12 hours, and carry out 24 hours at 25°C. For secondary drying within 1 hour, the lyophilized product was reconstituted with 2 mL of dis...

Embodiment 2

[0041] To the phosphate buffer (pH 7.5, concentration 50mmol / L) containing 10% trehalose, 200mM glycine, α-HBDH and LDH concentrations were respectively 400U / L and 550U / L, wherein: the concentration of phosphate buffer, Disodium hydrogen phosphate (Na 2 HPO 4 12H 2 O) 6.02g and sodium dihydrogen phosphate (NaH 2 PO 4 2H 2 (0) 0.5g is a benchmark, and the concentration after dissolving and constant volume in distilled water to 1000mL is 50mmol / L;

[0042]Add respectively according to the following formula: (1) No BSA (2) 1% BSA (2) 3% BSA (3) 5% BSA. Take 2 mL of the prepared solution of each group and put it into a brown vial for freeze-drying. Add 2 mL of distilled water to reconstitute the freeze-dried product and measure the activity of α-HBDH and LDH. Place the reserved freeze-dried product in a 37°C incubation The box was accelerated for 3 weeks and measured once a week. The results are shown in Table 2:

[0043] Table 2

[0044]

[0045] According to the resu...

Embodiment 3

[0047] To the phosphate buffer (pH value 7.5, concentration 50mmol / L) that contains 3%BSA, 200mM glycine, α-HBDH and LDH concentration are respectively 400U / L and 550U / L, wherein: the concentration of phosphate buffer, with Weigh disodium hydrogen phosphate (Na 2 HPO 4 12H 2 O) 6.02g and sodium dihydrogen phosphate (NaH 2 PO 4 2H 2 (0) 0.5g is a benchmark, and the concentration after dissolving and constant volume in distilled water to 1000mL is 50mmol / L;

[0048] Add respectively according to the following formula: (1) 1% trehalose (2) 6% trehalose (3) 12% trehalose (4) 16% trehalose (5) 20% trehalose. Take 2 mL of the prepared solution of each group and put it into a brown vial for freeze-drying. Add 2 mL of distilled water to reconstitute the freeze-dried product and measure the activity of α-HBDH and LDH. Place the reserved freeze-dried product in a 37°C incubation The box was accelerated for 4 weeks and measured once a week. The results are shown in Table 3:

[00...

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Abstract

The invention discloses a method for stabilizing activity of alpha-hydroxybutyricdehydrogenase and lactic dehydrogenase of quality-control serum, comprising the step of freezing and drying a mixture composed of other proteins, trehalose, mannite, glycine, quality-control serum and buffer solution. By the method in the invention, a freeze-drying quality-control serum containing alpha-HBDH and LDH and with stable properties and low price can be provided, the freeze-drying alpha-HBDH and LDH quality-control serum is stable and can be stored in a long term, the activity of the two enzymes of alpha-HBDH and LDH has no obvious change after hydrated reconstruction.

Description

technical field [0001] The invention relates to a method for stably quality-controlling the activity of serum α-hydroxybutyrate dehydrogenase or lactate dehydrogenase, the freeze-dried quality control containing α-hydroxybutyrate dehydrogenase or lactate dehydrogenase for clinical testing products or calibration products. Specifically, the present invention relates to a method for protecting the enzymatic activity of quality control substances in medical examinations, and in particular to quality control substances α-hydroxybutyrate dehydrogenase or lactate dehydrogenase after freeze-drying that do not show signs of A method in which the activity changes and can be preserved for a long time. technical background [0002] Lactate dehydrogenase (LDH for short, the same below) mainly exists in the heart, kidney, liver and muscle tissue. When these tissues are damaged, it will lead to the increase of LDH. The increase of LDH is mainly seen in myocardial infarction, hepatitis, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/96
Inventor 李子樵练子富崔鹏飞
Owner AILEX TECH GRP CO LTD
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