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Enzyme conjugate stabilizing solution

An enzyme conjugate and stabilizing solution technology, applied in the direction of enzyme stabilization, can solve problems such as easy loss of activity, and achieve the effects of non-toxic, harmless, easy preparation and simple operation for the human body

Active Publication Date: 2012-07-04
BIOSCIENCE (TIANJIN) DIAGNOSTIC TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In view of this, the present invention proposes an enzyme conjugate stabilizing solution to solve the problem that the enzyme conjugate in the related art is prone to lose activity after leaving the low-temperature storage environment

Method used

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Examples

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Embodiment 1

[0020] The TSH chemiluminescent immunoassay method established with the embodiment of the present invention to prepare anti-thyroid-stimulating hormone (TSH) monoclonal antibody-horseradish peroxidase (HRP) conjugate working solution includes the following steps:

[0021] Step 1, preparing enzyme conjugate stable solution, its component and concentration are:

[0022] 0.02mol / L phosphate (PBS) buffer system pH7.4

[0023] Casein 1g / L

[0024] Lactose 5g / L

[0025] Glycine 2g / L

[0026] Antipyrine 5g / L

[0027] Vitamin C 1g / L

[0028] Triton X-100 (Triton X-100) 0.2g / L

[0029] Gentamicin sulfate 500,000 units / L

[0030] Proclin 300 0.5g / L

[0031] Step 2. Coat the white light-emitting plate with anti-TSH monoclonal antibody, the coating concentration is 5ug / ml, 100ul / well, and coat at 37°C for 2 hours. After washing the plate, coat with blocking solution at 200 / well for 18 hours at 4°C Finally, discard the blocking solution in the coated plate, pat it dry, and dry it i...

Embodiment 2

[0039] The TSH chemiluminescent immunoassay method established with the embodiment of the present invention to prepare anti-thyroid-stimulating hormone (TSH) monoclonal antibody-horseradish peroxidase (HRP) conjugate working solution includes the following steps:

[0040] Step 1, preparing enzyme conjugate stable solution, its component and concentration are:

[0041] 0.02mol / L phosphate (PBS) buffer system pH7.4

[0042] Bovine Serum Albumin (BSA) 5g / L

[0043] Trehalose 25g / L

[0044] Tyrosine 10g / L

[0045] Antipyrine 20g / L

[0046] Vitamin C 2g / L

[0047] Tween-20 (Tween-20) 1g / L

[0048] Gentamicin sulfate 2 million units / L

[0049] Proclin 300 0.5g / L

[0050] Step 2. Coat the white light-emitting plate with anti-TSH monoclonal antibody, the coating concentration is 5ug / ml, 100ul / well, and coat at 37°C for 2 hours. After washing the plate, coat with blocking solution at 200 / well for 18 hours at 4°C Finally, discard the blocking solution in the coated plate, pat it...

Embodiment 3

[0058]The TSH chemiluminescent immunoassay method established with the embodiment of the present invention to prepare anti-thyroid-stimulating hormone (TSH) monoclonal antibody-horseradish peroxidase (HRP) conjugate working solution includes the following steps:

[0059] Step 1, preparing enzyme conjugate stable solution, its component and concentration are:

[0060] 0.02mol / L phosphate (PBS) buffer system pH7.4

[0061] Bovine Serum Albumin (BSA) 7.5g / L

[0062] Sucrose 50g / L

[0063] Lysine 30g / L

[0064] Antipyrine 50g / L

[0065] Vitamin C 10g / L

[0066] Tween-80 (Tween-80) 5g / L

[0067] Gentamicin sulfate 5 million units / L

[0068] Proclin 300 0.5g / L

[0069] Step 2. Coat the white light-emitting plate with anti-TSH monoclonal antibody, the coating concentration is 5ug / ml, 100ul / well, and coat at 37°C for 2 hours. After washing the plate, coat with blocking solution at 200 / well for 18 hours at 4°C Finally, discard the blocking solution in the coated plate, pat it d...

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Abstract

The invention provides an enzyme conjugate stabilizing solution which comprises the following components: 1-7.5g / L protein material, 5-50g / L saccharide material, 2-30g / L amino acid material, 5-50g / L antipyrine, 1-10 g / L vitamin C, 0.2-5g / L surfactant, 0.5-5 million IU / L gentamycin sulfate, 0.5g / L Proclin 300 and 0.02mol / L phosphate buffer solution the pH value of which is 7.4, wherein the protein material is bovine serum albumin or casein; the saccharide material is trehalose, gluecose, lactose or cane sugar; the amino acid material is glycine, phenylalanine, praline, tyrosine, serine, cysteine, methionine, glutamoyl or lysine; the surfactant is Twain-20, Twain-80 or triton x-100. The enzyme conjugate stabilizing solution provided by the invention solves the problem that the enzyme conjugate is easy to lose activity after the enzyme is not in the low temperature storage environment in related technology.

Description

technical field [0001] The invention relates to an enzyme conjugate stabilizing solution. Background technique [0002] At present, the most widely used immunoassay methods in the clinical field are enzyme-linked diagnostic reagents (EIA) and chemiluminescent immunoassays (CLIA). Among them, since the conjugates of trace antigens or antibodies and enzymes (HRP or AP) are easily inactivated after being formulated into ordinary buffers (such as PBS), both EIA and CLIA methods involve a key technology, that is, maintaining Stability of Enzyme Conjugates. [0003] The commonly used enzyme conjugate diluents cannot effectively maintain the activity of trace antigen or antibody and enzyme (HRP or AP) conjugates. The method to maintain the biological activity of the enzyme conjugate is usually to store the enzyme conjugate working solution in an environment below -20°C, and if it is stored at 2-8°C for one week or destroyed for one day at 37°C, its biological activity will be los...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/96
Inventor 刘萍栾大伟
Owner BIOSCIENCE (TIANJIN) DIAGNOSTIC TECH CO LTD
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