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Vascular endothelial growth factor (VEGF) receptor enzyme linked diagnosis kit and preparation method thereof

A detection kit and enzyme-linked technology, applied in the field of biomedicine, can solve the problems of low sensitivity and poor stability, and achieve the effect of broad application prospects, wide application scope and social needs, and effective monitoring means.

Inactive Publication Date: 2013-01-02
吴克
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The biological detection method for VEGF in the prior art has problems such as low sensitivity and poor stability

Method used

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  • Vascular endothelial growth factor (VEGF) receptor enzyme linked diagnosis kit and preparation method thereof
  • Vascular endothelial growth factor (VEGF) receptor enzyme linked diagnosis kit and preparation method thereof
  • Vascular endothelial growth factor (VEGF) receptor enzyme linked diagnosis kit and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1VE

[0030] Preparation of Example 1 VEGF receptor enzyme-linked diagnostic kit

[0031] The kit of the present invention can be exemplified by the following substances:

[0032] (1) VEGF receptor coated enzyme-linked plate;

[0033] (2) One positive control (lyophilized powder of VEGF protein);

[0034] (3) Negative control (animal serum freeze-dried powder) 1 tube;

[0035] (4) Sample diluent: 0.050r6 Tween 20, lor6 bovine serum albumin, 10mM edetate disodium, 0.050r6 thimerosal, 0.4M NaCl, pH6.5, 1 bottle,

[0036] (5) Enzyme conjugate: horseradish peroxidase-labeled vascular endothelial growth factor monoclonal antibody, 1 bottle;

[0037] (6) Concentrated washing solution: 137mM NaCl, 2.7mM KC1, 10mM Na2HP04, 2rrrMKH2P04, 0.050r6 Tween 20, Ph7.4, 1 bottle;

[0038] (7) Liquid level H of developer A. O. Solution, developer B solution is 3, 37, 5, 5 tetramethylbenzidine solution, 1 bottle each;

[0039] (8) The stop solution is 2MH, s04, 1 bottle;

[0040] (9) 2 sheets o...

Embodiment 2V

[0095] The usage method of embodiment 2VEFG enzyme-linked diagnostic kit

[0096] 1. Take out the required strips from the airtight bag equilibrated to room temperature, and seal the other strips and put them back at 4°C.

[0097] 2. Leave one well as a blank well, and add 100uL (or 2 drops) of sample diluent to the other wells.

[0098] 3. In addition to the blank well, add negative control 1 well, positive control 2 wells, and serum samples 100uL each to the corresponding wells, flick and mix well. Seal the reaction well with sealing tape and keep at 37°C for 60 minutes.

[0099] 4. Wash the plate 3 times: (1) Automatic plate washer: Shake off the liquid in the well, the washing liquid required to be injected is 350uL, the interval between injection and suction is 1530 seconds, and the plate is washed 3 times. (2) Manual plate washing: Shake off the liquid in the wells, add 350uL of washing solution to each well, let it stand for 30 seconds, shake off the liquid, pat d...

Embodiment 3

[0104] Example 3 Detection of kit linear range, linearity, specificity, sensitivity, accuracy, precision, repeatability

[0105] 1. Specificity

[0106] Specificity evaluation: The assembled VEGF receptor detection kit was used to detect the VEFG receptor, and the results showed that the specificity of the kit was high, as shown in Table 1.

[0107] Table 1 Result of 450nm absorbance of method specificity test for detecting VEGF receptor antigen content

[0108]

[0109]

[0110] The results show that the detection kit has high specificity and has no crossover with TNF-α, IL 22 and other receptors.

[0111] 2. Sensitivity

[0112] The detection is shown in Table 2 below:

[0113] Table 2 Sensitivity of VEGF receptor antigen detection methods

[0114]

[0115] The results show that: when the content is 1000-5ng / mL, the OD value ≥ negative control * 2.1, the detection sensitivity of this method is 5ng / mL detection range and linearity:

[0116] Table 3 VEGF receptor...

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Abstract

The invention discloses a vascular endothelial growth factor (VEGF) receptor enzyme linked diagnosis kit. A VEGF receptor coated enzyme linked reaction plate, an enzyme conjugate prepared through a horse radish peroxidase (HRP)-marked monoclonal antibody and a protecting agent, VEGF protein freeze-dried powder serving as positive comparison, protein freeze-dried powder serving as negative comparison, sample diluting liquid, PBST concentrated washing liquid, a chromogenic substrate prepared through 3,3',5,5'-tetramethyl benzidine, 2MH2S04 serving as stop buffer and shrouding adhesive paper are arranged in a kit body. The kit can be used for diagnosing and prognosis of diseases related to VEGF receptors such as cancer, angiopathy, diabetes retina disease and rheumatoid arthritis, has the advantages of being accurate, special, flexible, stable, convenient and the like, and has wide application prospects.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to an enzyme-linked detection kit for detecting human vascular endothelial cell growth factor (VEGF) and a preparation method thereof. Background technique [0002] Tumor development and metastasis are multi-stage, complex and highly selective processes involving the intricate relationship between tumor cells and hosts, and the mechanism is still unclear. In recent years, research on the relationship between new angiogenesis and tumor development, metastasis, and prognosis has progressed rapidly, proving that new angiogenesis is a necessary condition for tumorigenesis, metastasis, and the growth of metastatic foci, and vascular endothelial growth factor (VEGF) is the regulator of neovascularization. The most important cytokine produced. Studies have shown that vascular endothelial growth factor (VEGF) has an irreplaceable role in the development and differentiation ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/577G01N33/543
Inventor 吴克
Owner 吴克
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