Short peptide, detection kit and method for detecting vascular endothelial growth factor

A technology for detecting kits and growth factors, applied in the fields of biotechnology and immunoanalysis, can solve the problems of cumbersome operation process, low degree of automation, and cumbersome preparation process of the detection method, and achieve low cost, low commercialization cost, and good safety Effect

Active Publication Date: 2021-08-06
QINGDAO UNIV
View PDF6 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The inventors of the present invention found that although antibodies and aptamers have good selectivity as capture probes, their inherent weaknesses (such as cumbersome preparation process, low stability and high cost) limit their application, In addition, the existing detection methods are not only cumbersome to operate, but also have a low degree of automation and are greatly affected by human factors.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Short peptide, detection kit and method for detecting vascular endothelial growth factor
  • Short peptide, detection kit and method for detecting vascular endothelial growth factor
  • Short peptide, detection kit and method for detecting vascular endothelial growth factor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment approach

[0026] A typical embodiment of the present invention provides a short peptide, which is polypeptide 1 or polypeptide 2;

[0027] The polypeptide 1 is: a polypeptide consisting of an amino acid sequence of LSTSPHTGGGSC;

[0028] The polypeptide 2 is: a polypeptide derived from the polypeptide 1 through substitution and / or deletion and / or addition of one or several amino acid residues and having the same function.

[0029] It was found through experiments that the short peptide not only has the advantages of high affinity and high selectivity for VEGF165, but also has the advantages of small molecular weight and good stability. The short peptide is a short-chain polypeptide. The short peptide is moderately extended while ensuring the binding site, so as to enhance the binding efficiency with the polystyrene surface, and at the same time facilitate more sufficient exposure of the probe binding site.

[0030] Another embodiment of the present invention provides a detection kit, ...

Embodiment 1

[0045] Probe V for specificity testing:

[0046] Dissolve the specific synthetic probe V in 0.1M TBS (pH 7.4) to a concentration of 10 μg / mL, then load 100 μL of the prepared solution into the wells of a 96-well microtiter plate, and store the sealed plate at 4 °C Incubate overnight; the next day, suck out the synthetic peptide probe solution in the ELISA plate, place the plate face down on a clean paper towel, pat dry the residue, add 0.5% TBST to rinse: 200 μL, 6 times, 2 min each time; antigen ( 5ng / mL, 100μL): vascular endothelial growth factor 165 (VEGF165), vascular endothelial growth factor 121 (VEGF121), human immunoglobulin G (HIgG), human immunoglobulin A (HIgA), human serum albumin (HSA) , prostate specific antigen (PSA), carbohydrate antigen 15-3 (CA15-3), cancer antigen 19-9 (CA19-9), carbohydrate antigen 242 (CA242) and carcinoembryonic antigen (CEA) were placed in 96 After incubating the well plate at room temperature for 3 hours, wash 6 times with TBST and pat...

Embodiment 2

[0049] Probe V optimal coating concentration test:

[0050] Serial serial dilutions of specific probe V: 2, 5, 10, 20, 30, 40 μg / mL were added to the wells of a 96-well white microplate, 100 μL per well, wrapped with plastic wrap, and placed in 4 Incubate at ℃ overnight with shaking; suck out the synthetic peptide probe solution in the ELISA plate, place the plate face down on a clean paper towel, pat dry the residue, add 0.5% TBST to rinse: 200 μL per well, wash 6 times, 2 min each time ; Add 1 μg / mL VEGF165 100 μL to the well one by one, incubate with shaking at room temperature for 3 h; wash 6 times with TBST and pat dry; dilute the anti-VEGF165 polyclonal antibody in the buffer to a concentration of 1:1000, add 100 μL to the well And shake and incubate at room temperature for 1 hour, then wash 6 times with TBST and pat dry; Dilute the IgG-HRP monoclonal antibody in the buffer to a concentration of 1:500, add 100 μL of the diluted conjugate to each well, And incubated at r...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention discloses a short peptide, a detection kit and a method for detecting vascular endothelial growth factor. The short peptide is polypeptide 1 or polypeptide 2; the polypeptide 1 is a polypeptide composed of an amino acid sequence of LSTSPHTGGGSC; and the polypeptide 2 is a polypeptide derived from the polypeptide 1 that has been substituted and / or deleted and / or added to one or several amino acid residues and has the same function. The short peptide can be used as a capture antibody and to detect a specified tumor marker in serum combined with a chemiluminescent enzyme immunoassay (CLIA). The solid-phase biological polypeptide probe and target protein in a serum sample to be detected are incubated, and then effective detection can be realized under a catalytic color development system. The method provided by the invention is strong in specificity, high in sensitivity and convenient to operate, has relatively low probe production and purification cost, and provides an effective way for realizing sensitive and rapid detection of VEGF165.

Description

technical field [0001] The invention belongs to the technical field of biotechnology and immune analysis, and relates to a short peptide, a detection kit and a method for detecting vascular endothelial growth factor. Background technique [0002] The information disclosed in this background section is only intended to increase the understanding of the general background of the present invention, and is not necessarily taken as an acknowledgment or any form of suggestion that the information constitutes the prior art already known to those skilled in the art. [0003] Vascular endothelial growth factor (VEGF) is a highly glycosylated basic protein with an isoelectric point of 8.5 and a molecular weight of 45KD. It is formed by combining two identical subunits through disulfide bonds and has strong acid and heat resistance. VEGF is an extremely important cytokine that regulates neovascularization. There are five different subtypes of VEGF, namely VEGF165, VEGF189, VEGF206, VE...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/08G01N33/68G01N33/577
CPCC07K7/08G01N33/68G01N33/577G01N2333/515G01N2410/00
Inventor 刘爱骅王戈
Owner QINGDAO UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap