Enzymatic detection kit for small and dense low density lipo-protein cholesterol

A low-density lipoprotein and detection kit technology, applied in the field of biomedicine, can solve the problems of incomplete precipitation, inaccurate measurement results, and inapplicability to automatic biochemical analyzers, etc., to achieve simple operation, good specificity, and high measurement results Accurate and reliable results

Pending Publication Date: 2020-12-04
上海睿康生物科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method does not require special instruments and equipment, and is simpler than the traditional detection method of LDL subcomponents; but it has the disadvantage that pre-precipitation treatment is prone to incomplete precipitation, which may easily

Method used

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  • Enzymatic detection kit for small and dense low density lipo-protein cholesterol
  • Enzymatic detection kit for small and dense low density lipo-protein cholesterol
  • Enzymatic detection kit for small and dense low density lipo-protein cholesterol

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] The basic flow of embodiment 1 prokaryotic expression system

[0033] Obtain the target gene, use the cloning plasmid containing the target gene as a template by PCR method, design a pair of primers according to the gene sequence, introduce different enzyme cutting sites into the upstream and downstream primers, and obtain the desired gene fragment by PCR cycle.

[0034]It should be noted that monoclonal antibody 1 and monoclonal antibody 2 are designed for specific sites of peptide 82-192 and peptide 202-35 of high-density lipoprotein respectively, and monoclonal antibodies can be prepared using hybridoma technology, for example The monoclonal antibody is produced by a hybridoma cell line selected from the following group: CCTCC-C200923, CCTCC-C200924 or CCTCC-C200925 (specific preparation methods can be found in: Ou Lanxiang et al., International Journal of Laboratory Medicine, 2019, 40 (22), recombinant Prokaryotic expression of human lipoprotein-associated phospholi...

Embodiment 2

[0039] In this example, the interference effect of anti-human high-density lipoprotein antibody on eliminating high-density lipoprotein was evaluated, and the scheme is as follows, see Table 1:

[0040] Table 1.

[0041]

[0042]

[0043] Table 2 is the specificity experimental evaluation. Wherein, the experimental condition parameters adopted in Table 2 are as follows:

[0044] Validation reagent batch number: 20190601

[0045] calibrator

[0046] Batch number of the company's supporting calibration products: 20190601

[0047] Machines used and parameter settings:

[0048] Dirui CS-600B automatic biochemical analyzer, the parameters are set as follows

[0049] SDLDL: 660nm / 546nm; two-point endpoint method; 3 / 270 / 90

[0050] Table 2.

[0051]

[0052] Therefore, it can be seen from Table 1 and Table 2 that the addition of anti-human high-density lipoprotein antibody (as shown in Scheme 2-4) has a significant effect on inhibiting high-density lipoprotein, and th...

Embodiment 3

[0054] This implementation case mainly evaluates the concentrations of the two antibodies used. The scheme is shown in Table 3 below:

[0055] table 3.

[0056]

[0057] Table 4 shows the specificity experimental evaluation of the protocol used in Table 3.

[0058] Table 4.

[0059]

[0060] Conclusion: The higher the concentration of the antibody used, the higher the inhibition rate of high-density lipoprotein. When it reaches 12mg / L, the inhibition rate reaches the best

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Abstract

The invention discloses an enzymatic detection kit for small and dense low density lipo-protein cholesterol. The enzymatic detection kit for the small and dense low density lipo-protein cholesterol comprises a reagent R1, a reagent R2 and a calibrator; the reagent R1 consists of the following components at specific concentrations: a buffer solution, cholesterol lipase, cholesterol oxidase, catalase, an anti-human high-density lipo-protein antibody, a surfactant, and a preservative; the reagent R2 consists of the following components at specific concentrations: a buffer solution, a surfactant,4-aminoantipyrine, peroxidase, and a preservative; and the calibrator consists of the following components at specific concentrations: a buffer solution, glycerol, a stabilizer, a small and dense lowdensity lipo-protein cholesterol (sdLDL) antibody, and a preservative. In virtue of addition of the anti-human high-density lipo-protein antibody, the enzymatic detection kit for the small and dense low density lipo-protein cholesterol has greatly improved kit performance, better reagent stability, higher sensitivity, wider detection range and reduced detection cost.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a protease detection kit, in particular to a small and dense low-density lipoprotein cholesterol enzymatic detection kit. Background technique [0002] The Lipoprint LDL Subfraction Rapid Analysis System is currently the only diagnostic device certified by the US Food and Drug Administration for the separation and detection of LDL subfractions. The Lipoprint detection system is based on the particle size and charge of LDL to achieve the purpose of separation through polyacrylamide gel electrophoresis. LDL can be divided into 7 subcomponents in a short time, of which components 1 and 2 are defined as ldLDL particles; fractions 3 to 7 are defined as sdLDL particles. The system can efficiently detect subcomponents of LDL quantitatively and qualitatively, and thus is more conducive to popularization and application in clinical routine laboratories. Studies have shown that in the plasma o...

Claims

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Application Information

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IPC IPC(8): C12Q1/60C12Q1/44C12Q1/30C12Q1/28C12Q1/26
CPCC12Q1/60C12Q1/44C12Q1/30C12Q1/28C12Q1/26G01N2333/904C12Q2326/96
Inventor 房君江赵猛
Owner 上海睿康生物科技有限公司
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