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Kit as well as preparation method and application thereof

A technology of kits and reagents, applied in biological testing, analysis through chemical reactions of materials, instruments, etc., can solve problems such as poor anti-interference ability, large test deviation, and susceptibility to environmental influences

Pending Publication Date: 2022-05-27
深圳天辰医疗科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Immunoturbidimetry This method has high requirements for samples and poor anti-interference ability. For hemolysis samples and blood lipid samples, there will be large test deviations, and the sensitivity is relatively low. Fluorescence immunochromatography is simple, efficient and easy to operate, and does not require large instruments. However, due to the limitation of the membrane medium, there are defects such as large test deviation, easy to be affected by the environment, and low sensitivity.

Method used

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  • Kit as well as preparation method and application thereof
  • Kit as well as preparation method and application thereof
  • Kit as well as preparation method and application thereof

Examples

Experimental program
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preparation example Construction

[0070] The present invention further proposes a preparation method of the kit as described above, comprising the following steps:

[0071] S10. Streptavidin magnetic beads are added to the first diluent, mixed evenly, washed by magnetic separation, and then resuspended in the first diluent to obtain the first reagent.

specific Embodiment approach

[0073]Take 10 mg streptavidin magnetic beads, add 1 mL of the first dilution (50 mM 2-morpholineethanesulfonic acid, 0.05% Tween 20 (v / v), 0.1% Proclin300 (v / v), 0.9% NaCl ( w / v), 1% bovine serum albumin (w / v), 0.1% glycine (w / v), 0.5% trehalose (w / v) and 0.05% BSA-modified PEG (w / v), pH value of 6.5), magnetic separation and washing, repeated washing 3 times, resuspended in the first diluent, and diluted with the first diluent so that the concentration of streptavidin magnetic beads is 0.05mg / mL~0.2mg / mL, that is as the first reagent.

[0074] S20. After the soluble fms-like tyrosine kinase-1 monoclonal antibody is activated, mixed with the enzyme solution for reaction and purified, the enzyme-labeled soluble fms-like tyrosine kinase-1 monoclonal antibody is prepared, and the enzyme-labeled soluble The fms-like tyrosine kinase-1 monoclonal antibody is uniformly mixed with the second diluent to obtain the second reagent.

[0075] Specifically, in the embodiment of the presen...

Embodiment 1

[0100] Example 1 Kit

[0101] The first reagent: streptavidin magnetic beads and the first diluent [50mM 2-morpholineethanesulfonic acid, 0.05% Tween 20 (v / v), 0.1% Proclin300 (v / v), 0.9% NaCl ( w / v), 1% bovine serum albumin (w / v), 0.1% glycine (w / v), 0.5% trehalose (w / v) and 0.05% BSA-modified PEG (w / v), pH Value is 6.5], the concentration of streptavidin magnetic beads is 0.05mg / mL, and the particle diameter of streptavidin magnetic beads is 1.5 μm;

[0102] Second reagent: enzyme-labeled soluble fms-like tyrosine kinase-1 monoclonal antibody and second diluent [50mM 2-morpholineethanesulfonic acid, 0.05% Tween 20 (v / v), 0.1% Proclin300 (v / v) v), 0.9% NaCl (w / v) and 1.5% bovine serum albumin (w / v), the pH value is 6.8], and the enzyme is alkaline phosphatase;

[0103] The third reagent: NHS-LC-biotin labeled with soluble fms-like tyrosine kinase-1 monoclonal antibody and the third dilution [50mM 2-morpholineethanesulfonic acid, 0.05% Tween 20 (v / v), 0.1% Proclin300 (v / v),...

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Abstract

The invention discloses a kit and a preparation method and application thereof, and relates to the technical field of immunodetection analysis, the kit is used for detecting the content of soluble fms-like tyrosine kinase-1, the kit comprises: a first reagent comprising streptavidin magnetic beads and a first diluent; the second reagent comprises an enzyme-labeled soluble fms-like tyrosine kinase-1 monoclonal antibody and a second diluent; the third reagent comprises NHS-LC-biotin marked with a soluble fms-like tyrosine kinase-1 monoclonal antibody and a third diluent; a calibrator; and a light-emitting substrate. When the kit provided by the invention is used for detecting the content of soluble fms-like tyrosine kinase-1, the sensitivity is high, the stability and the repeatability are good, and the anti-interference capability is strong.

Description

technical field [0001] The invention relates to the technical field of immunoassay and analysis, in particular to a kit and its preparation method and application. Background technique [0002] sFlt-1 (soluble fms-like tyrosine kinase-1) is a rare spliced ​​form of vascular endothelial growth receptor-1 (fms-like tyrosinekinase receptor 1, Flt-1), which only retains the ligand-binding domain and does not have Tyrosine kinase activity, molecular weight 63.6KD, produced by trophoblast cells during pregnancy. Medical research at home and abroad has shown that sFlt-1 inhibits angiogenesis by affecting the effects of pro-angiogenic factors VEGF (vascular endothelial growth factor) and PIGF (placental growth factor); the expression of VEGF and PIGF in the placenta of PE (preeclampsia) patients is reduced , and deduced that the decreased expression of VEGF family proteins affected the differentiation and invasion of placental trophoblast cells, and even resulted in placental hypox...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/577G01N33/543G01N33/58G01N21/76
CPCG01N33/6893G01N33/74G01N33/577G01N33/54326G01N33/581G01N21/76G01N2800/368G01N2333/71
Inventor 邓家兵黄金浪唐灿
Owner 深圳天辰医疗科技有限公司
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