Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Fluorescence immunochromatographic test paper for detecting human lp-pla2 protein and preparation method thereof

A fluorescence immunochromatography, lp-pla2 technology, applied in biological testing, measuring devices, analytical materials, etc., to achieve the effects of optimized preparation conditions, wide detection range, and improved fluorescence signal-to-background ratio

Active Publication Date: 2016-05-18
深圳市安群生物工程有限公司
View PDF7 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is currently no fluorescent immunochromatographic test strip for human Lp-PLA2 protein

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
  • Fluorescence immunochromatographic test paper for detecting human lp-pla2 protein and preparation method thereof
  • Fluorescence immunochromatographic test paper for detecting human lp-pla2 protein and preparation method thereof
  • Fluorescence immunochromatographic test paper for detecting human lp-pla2 protein and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0111] Example 1: Preparation of Lp-PLA2 epitope peptides (1) and (2).

[0112]The preparation method uses chemical synthesis method: Lp-PLA2 antigenic epitope peptides (1) and (2) are synthesized respectively by solid-phase method using the American ABI431A automatic peptide synthesizer. The purity of the epitope peptide was assessed by high performance liquid chromatography, and the concentration of the peptide was determined. The molecular weights of the antigenic epitope peptides (1) and (2) of the present invention are 1690.09 and 1623.92 respectively, which are determined by mass spectrometry, and the synthesized polypeptide sequences are identified by polypeptide sequence determination.

[0113] 1. Synthesis of Lp-PLA2 epitope peptides (1) and (2)

[0114] The above peptides were synthesized by solid-phase method. The main idea of ​​solid-phase peptide synthesis is: first connect the carboxyl group of the carboxyl-terminal amino acid of the peptide chain to be synthes...

Embodiment 2

[0202] Embodiment 2: the Lp-PLA2 antigen epitope peptide (1) and (2) obtained in Example 1 are linked with carrier protein to prepare Lp-PLA2 antigen (1) and (2), utilize gained antigen (1) and (2) Animals are immunized separately to prepare specific monoclonal antibodies and polyclonal antibodies using the antigen (1), and specific monoclonal antibodies and polyclonal antibodies are prepared using the antigen (2).

[0203] 1. Antigen preparation: Lp-PLA2 peptides (1) and (2) were linked with carrier protein KLH (keyhole limpet hemocyanin) by BDB (Bis-diazotizedbenzidinedichloride) method to prepare Lp-PLA2 antigens (1) and (2).

[0204] Take 10.0mg of Lp-PLA2 peptide (1) or (2), dissolve it with 1ml0.1MPBS buffer (pH7.4); dissolve KLH10mg with 20ml of 0.2M borate buffer (pH9.0); Mix them, cool to 0°C, take BDBCl 2 110 μL, reacted at room temperature for 1.5 h, dialyzed overnight, then aliquoted, and stored at -20 °C.

[0205] In the present embodiment, the formula of PBS d...

Embodiment 3

[0219] Example 3: Specific identification of human Lp-PLA2 monoclonal antibodies (1) and (2)

[0220] Detection was performed by ELISA. Human Lp-PLA2 protein, fibrinogen, and C-reactive protein (all purchased from Shanghai Lianshuo Company) were used as detection antigens to coat ELISA plates, and the prepared Lp-PLA2 monoclonal antibodies (1) and (2) For the specific reaction with the human Lp-PLA2 protein, normal BALB / c mouse serum was used as negative control, and PBS solution was used as blank control.

[0221] Result: Lp-PLA2 monoclonal antibody (1) and (2) only react positively (P / N>2.1) with Lp-PLA2 respectively, and are negative with fibrinogen, C-reactive protein reaction, illustrate that the present invention Lp-PLA2 monoclonal antibodies (1) and (2) have specificity respectively.

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

PropertyMeasurementUnit
particle diameteraaaaaaaaaa
wavelengthaaaaaaaaaa
wavelengthaaaaaaaaaa
Login to View More

Abstract

The invention relates to fluorescence immunochromatography test paper for detecting human Lp-PLA2 proteins and a preparation method of the fluorescence immunochromatography test paper. The test paper is used for detecting the human Lp-PLA2 proteins by virtue of a two-antibody sandwich method, and in the two-antibody sandwich method, a first Lp-PLA2 monoclonal antibody which is labeled with fluorescence microspheres serves as a trapping antibody, and is sourced from one of sequences indicated in sequence tables of SEQ ID NO.1 and SEQ ID NO.2; a second Lp-PLA2 monoclonal antibody serves as a detection antibody, and is sourced from the other one of the sequences indicated in the sequence tables of SEQ ID NO.1 and SEQ ID NO.2. The fluorescence immunochromatography test paper is simple and rapid to operate, wide in detection range, high in specificity and good in sensitivity.

Description

technical field [0001] The invention belongs to the field of polypeptide chemistry and immunology, and in particular relates to a human lipoprotein-associated phospholipase A2 (Lp-PLA2) epitope peptide, an Lp-PLA2 specific antigen prepared by using the epitope peptide, and a corresponding monoclonal antibody or Polyclonal antibody, use of said antibody in preparing human Lp-PLA2 in vitro diagnostic kit, human Lp-PLA2 in vitro diagnostic kit, and a fluorescent immune layer for quantitative detection of human Lp-PLA2 protein in analyte Analysis test paper and its preparation method. Background technique [0002] Cardiovascular and cerebrovascular diseases have always been recognized as one of the biggest killers of human health and the leading cause of death worldwide. The number of people who die from cardiovascular and cerebrovascular diseases in my country is as high as 3 million or more every year, and there are more than 60 million patients. In recent years, with the co...

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 Patents(China)
IPC IPC(8): G01N33/577
CPCG01N33/566G01N33/92
Inventor 朱建安
Owner 深圳市安群生物工程有限公司
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com