Apolipoprotein E testing reagent

A technology for detecting reagents and apolipoproteins, which is applied in the field of reagents for measuring APOE in serum, can solve problems such as large differences in measurement results, poor stability, and inconvenient operation, and achieve simplification of operation methods and steps, improvement of anti-interference ability, Effect of detection efficiency improvement

Inactive Publication Date: 2010-08-11
浙江伊利康生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Known methods for measuring APOE include chromatography, electrophoresis, and immunoassay. Among them, chromatography and electrophoresis are cumbersome to operate, and they cannot perform batch sample analysis and directly go to a fully automatic biochemical analyzer. Immunodiffusion, radioimmunoassay, fluorescence-labeled immunoassay, enzyme-labeled immunoassay, and chemiluminescence immunoassay also have many deficiencies; if special equipment is required, the sample needs to be pretreated, and it cannot be used for automatic biochemical analysis. instrument for batch detection and analysis, etc.
The immunoturbidimetric method commonly used in clinical practice is popular because of its small amount of sample, its ability to directly perform batch sample analysis on a fully automatic machine, and its simple operation; however, the currently established methods and reagents have some shortcomings and / or deficiencies. The manifestations are: first, the antibody titer is not high, and the specificity, affinity, and affinity are not good; second, the calibration serum is based on human serum, although the HBsAg negative, HIV test negative, and alanine aminotransferase normal Human serum, but it is difficult to rule out the possibility of no other infectious diseases, which brings great danger to the operator; moreover, the calibration serum is a freeze-dried product, and the value of each batch is different, so it needs to be reconstituted with water before use. The operation is extremely inconvenient; in addition, there are great differences between different reconstituted bottles after reconstitution. After reconstitution, it must be frozen and stored, especially not repeatedly thawed for use. , and its stability is not good; third, the anti-interference ability of high-fat, jaundice, and hemolytic samples is poor; High phenomenon, the measured results are inaccurate; fifth, the antiserum precipitates in a short period of time, making it difficult to operate on the machine and affecting the detection effect

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Reagent R1 (reactant):

[0035] Phosphate (buffer) 25mmol / L (5-200mmol / L)

[0036] Polyoxyethylene lauryl ether (surfactant) 5% (0.1-10%)

[0037] NaCl (electrolyte) 4.5% (0.1%-0.5%)

[0038] PEG-6000 (polymer accelerator) 40% (20%-80%)

[0039] Broad-spectrum fungicide (preservative) 3% (0.1%-5%)

[0040] Polybrene (reaction accelerator) 0.3% (0.1%-0.5%)

[0041] Disodium EDTA (stabilizer) 0.5% (0.1%-1%)

[0042] Bovine serum albumin (stabilizer) 1% (0.1%-1%)

[0043] APOE antibody reagent

[0044] The rest is purified water

[0045] Reagent R2 (anti-APOE antibody solution):

[0046] Reagent diluent (or sample diluent) 1000ml

[0047] Goat anti-human APOE antiserum 400ml

[0048] Butylated hydroxyanisole 0.03%

[0049] Constant value calibration solution (serotype solution):

[0050] CAPSO (buffer) 20mmol / L

[0051] Butylated hydroxyanisole (antioxidant) 6mmol / L

[0052] Citric acid (synergist) 10mmol / L

[0053] Broad-spectrum fungicide (preservative) 2mmo...

Embodiment 2

[0061] Embodiment 2 Reagent R1 (reactant)

[0062] Phosphate (buffer) 30mmol / L

[0063] Polyoxyethylene lauryl ether (surfactant) 5%

[0064] NaCl (electrolyte) 4.5%

[0065] PEG-6000 (polymer accelerator) 40%

[0066] Broad-spectrum fungicide (preservative) 3%

[0067] Polybrene (reaction accelerator) 0.3%

[0068] Disodium edetate (stabilizer) 0.5%

[0069] Bovine serum albumin (stabilizer) 1%

[0070] The rest is purified water

[0071] Reagent R2 (anti-APOE antibody solution)

[0072] Reagent diluent (sample diluent) 1000ml

[0073] Goat anti-human APOE antiserum 250ml

[0074] Butylated hydroxyanisole 0.03%

[0075] Constant value calibration solution (serotype solution)

[0076] CAPSO (buffer) 50mmol / L

[0077] Butylated hydroxyanisole (antioxidant) 6mmol / L

[0078] Citric acid (synergist) 10mmol / L

[0079] Broad-spectrum fungicide (preservative) 2mmol / L

[0080] Disodium EDTA (stabilizer) 2mmol / L

[0081] Bovine serum albumin (stabilizer) 3mmol / L

[008...

Embodiment 3

[0088] Reagent R1 (reactant)

[0089] Phosphate (buffer) 200mmol / L (5-200mmol / L)

[0090] Polyoxyethylene lauryl ether (surfactant) 5% (0.1-10%)

[0091] NaCl (electrolyte) 4.5% (0.1%-0.5%)

[0092] PEG-6000 (polymer accelerator) 40% (20%-80%)

[0093] Broad-spectrum fungicide (preservative) 3% (0.1%-5%)

[0094] Polybrene (reaction accelerator) 0.3% (0.1%-0.5%)

[0095] Disodium EDTA (stabilizer) 0.5% (0.1%-1%)

[0096] Bovine serum albumin (stabilizer) 1% (0.1%-1%)

[0097] The rest is purified water

[0098] Reagent R2 (anti-APOE antibody solution)

[0099] Reagent (sample diluent) 1000ml

[0100] Goat anti-human APOE antiserum 300ml

[0101] Butylated hydroxyanisole 0.03%

[0102] Constant value calibration solution (serotype solution)

[0103] CAPSO buffer 100mmol / L

[0104] Butylated hydroxyanisole antioxidant 6mmol / L

[0105] Citric acid synergist 10mmol / L

[0106] Broad-spectrum fungicide preservative 2mmol / L

[0107] EDTA disodium stabilizer 2mmol / L

...

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Abstract

The invention relates to a reagent for determining APOE in blood serum, aiming to provide a stable reagent which has simple operation, high accuracy, good repeatability and strong anti-jamming capability, uses excellent new-born calf serum to replace human blood serum as substrate preparation and uses liquid blood serum type constant value calibration solution to detect apolipoprotein E in combination, wherein the sample does not need thinning dilution. The reagent is characterized in that the apolipoprotein E testing reagent comprises: a. a reagent R1 comprising buffer solution, a surface active agent, an electrolyte, a polymer accelerant, a reaction promoter, a stabilizer, a proper amount of preservative, and the balance of purified water; b. a reagent R2, wherein a certain amount of the reagent R1 is added to anti-human-APOE antiserum and antioxidant with the vacuum ratio of 20-50%; and c. constant value calibration solution comprising buffer solution, APOE antigen, antioxidant, synergist, stabilizer, proper amount of preservative, and the balance of purified water.

Description

technical field [0001] The invention relates to a preparation method and a reagent kit for measuring serum components, in particular to a reagent for measuring APOE in serum, which can be widely used in the technical fields of medicine and biochemistry. Background technique [0002] Apolipoprotein E (APOE) is a glycoprotein containing 299 amino acids combined with phospholipids. Its molecular weight is 34kD. ApoE can be synthesized in various tissues, but mainly in the liver. First, a propeptide containing 317 amino acids is synthesized. Mature ApoE is formed by sylation and desialylation of extracellular fluid; ApoE is transferred into lipoproteins after being secreted into the blood. APOE is a structural protein necessary for lipoproteins. It can participate in lipoprotein metabolism, regulate cholesterol levels, activate lecithin sterol lipid transferase (LCAT) by binding to the receptors of various tissue cells, and has a certain Immunomodulatory effect. Determination...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/539G01N33/96G01N21/31
Inventor 王贤理蒙凯蔡其浩
Owner 浙江伊利康生物技术有限公司
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