ELISA kit for lipoprotein A and preparation method thereof

An enzyme-linked immunoassay and lipoprotein technology, applied in the field of medical detection, can solve the problems of long detection time, low sensitivity, and complicated operation

Active Publication Date: 2018-03-02
ANHUI IPROCOM BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, the known methods for detecting the concentration of lipoprotein (a) include immunoturbidimetry, radioimmunoassay, fluorescent immunoassay and other detection methods, but the operation of the above-mentioned determination methods is relatively complicated, and the detection time is long or the sensitivity is low. , not suitable for routine testing

Method used

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  • ELISA kit for lipoprotein A and preparation method thereof
  • ELISA kit for lipoprotein A and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] (1) Preparation of a microtiter plate coated with anti-lipoprotein a antibody:

[0033] A, antibody dilution: the anti-lipoprotein a monoclonal antibody is diluted to 10 μ g / ml with the Tris-HCl buffer solution of 50 mM that pH is 8.0, obtains coating solution;

[0034] b. Coating: Take the microwell plate, wash it 3 times with washing solution, add the above-mentioned coating solution containing anti-lipoprotein a monoclonal antibody, 100 μL / well per well, and incubate at 4° C. for 12 hours;

[0035] c, sealing: incline coating liquid, be placed on absorbent paper and pat several times, remove raffinate, add the concentration that is 0.1% glycerol monoricinoleic acid ester, 0.5% BSA and 1% sucrose containing percentage by weight. 50mmol / L Tris-HCl blocking solution, pH 8.0, 300μl / well, room temperature, 1 hour;

[0036] d. Vacuum drying and sealing to obtain an ELISA plate coated with anti-lipoprotein a antibody.

[0037] (2) Preparation of enzyme-labeled antibody so...

Embodiment 2

[0055] Embodiment 2 kit sensitivity measurement

[0056] Prepare PBS buffer solution with different concentrations of lipoprotein a standard product respectively, the concentrations are respectively 0.1, 0.2, 0.5, 1 and 2 mg / L, and the kit prepared in Example 1 is used for detection, and the control buffer solution is used as a blank control, and the specific detection Methods as below:

[0057] a) Antigen-antibody reaction: Add 50 μl of standard solution and diluent to the microwells of the coated microtiter plate, and incubate in a water bath at 37° C. for 50 minutes. Wash the plate with washing buffer 5 times.

[0058] b) Add HRP-labeled anti-lipoprotein a antibody solution to each well, 100 μl per well, and incubate in a 37° C. water bath for 50 minutes. Repeat the plate washing operation 5 times.

[0059] c) Chromogenic reaction: each well was sequentially added with substrate solution, 50 μl each of the chromogenic solution, incubated in a water bath at 37° C. for 20 ...

Embodiment 3

[0065] Embodiment 3 test kit stability investigation

[0066] After the kit prepared in Example 1 was placed at 20°C for 6 months and 12 months respectively, the sensitivity of the kit and the absorbance of each concentration of the standard were measured according to the method of Example 2, and regression analysis was performed on the data to calculate R 2 value.

[0067] In this embodiment, the comparison ratio is set as follows:

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Abstract

The invention belongs to the field of medical assay, and in particular relates to an enzyme linked immunosorbent assay kit of lipoprotein a. The kit comprises the following components: (1) an enzyme label plate coated with an anti-lipoprotein a antibody; (2) lipoprotein a series standard products; (3) an enzyme label antibody solution; (4) diluent; (5) washing liquid; (6) a substrate solution; (7) a developing solution; and (8) a stop solution. The enzyme label antibody solution is a 50 mmol / L PBS buffer solution containing 5 mg / L HRP label anti-lipoprotein a antibody, 0.5 mg / L glycerinum single ricinoleate and 0.5 mg / L BSA, and the pH value is 8.0.

Description

technical field [0001] The invention belongs to the field of medical detection, and in particular relates to an ELISA kit for detecting phosphorylated lipoprotein (a) and a preparation method thereof. Background technique [0002] Atherosclerosis (atherosclerosis AS) refers to the lesion with thickening, hardening and decreased elasticity of the arterial wall, and is characterized by the formation of atherosclerotic plaques in the arterial intima. At present, the cause of the disease has not been fully determined, and it may be related to factors such as age, gender, dyslipidemia, hypertension, smoking, diabetes, obesity, and infection. High cholesterol concentration, lipoprotein a is a major risk factor for AS. [0003] Lipoprotein (a) is a special class of lipoprotein particles in human plasma, first discovered by Kare Berg in 1963, mainly composed of low-density lipoprotein particles and highly hydrophilic apo(a) rich in sugar chains; Each apo(a) molecule is covalently ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68G01N33/531
CPCG01N33/531G01N33/6893
Inventor 陈立国邹伟权张亚丽李庆祥母润红王涛苏焱
Owner ANHUI IPROCOM BIOTECH CO LTD
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