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

C-peptide monoclonal antibody cross-linked with magnetic particles, preparation method thereof and C-peptide test kit including same

A monoclonal antibody and detection kit technology, applied in the field of in vitro diagnostic medical testing, can solve the problems of poor stability of test results, weak anti-interference, and insufficient sensitivity

Active Publication Date: 2014-12-17
SHANGHAI KEHUA BIO ENG
View PDF5 Cites 4 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, the diluent for diluting the C-peptidase reagent is required in the currently used C-peptide detection kit, but the diluent also affects the stability of the C-peptidase reagent
At present, the C-peptide detection kits in the prior art have disadvantages such as non-specific binding, insufficient sensitivity, weak anti-interference, and poor stability of detection results.
The existing magnetic particles are directly washed and blocked with some irrelevant proteins (such as BSA) after cross-linking, but there are batch-to-batch differences in BSA, and the specificity of different BSAs is also different. Direct blocking with BSA often cannot fully achieve the ideal. The closing effect of

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
  • C-peptide monoclonal antibody cross-linked with magnetic particles, preparation method thereof and C-peptide test kit including same
  • C-peptide monoclonal antibody cross-linked with magnetic particles, preparation method thereof and C-peptide test kit including same
  • C-peptide monoclonal antibody cross-linked with magnetic particles, preparation method thereof and C-peptide test kit including same

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0059] Correspondingly, the present invention also provides a method for preparing the above-mentioned C-peptide monoclonal antibody cross-linked magnetic particles, and the method includes the following steps:

[0060] a) Cross-link the C-peptide monoclonal antibody with magnetic particles; and

[0061] b) Pretreating the C-peptide monoclonal antibody cross-linked magnetic particles obtained in step a) with a non-ionic surfactant.

[0062] The C-peptide monoclonal antibody cross-linked magnetic particles of the present invention can be used to detect C-peptide or prepare a C-peptide detection kit, thereby significantly improving the sensitivity in detecting C-peptide.

[0063] The term "nonionic surfactant" as used herein has the meaning commonly understood by those of ordinary skill in the art, that is, a surfactant that does not generate ions in an aqueous solution. Non-ionic surfactants do not dissociate when dissolved in water. The lipophilic group in the molecule is roughly the ...

Embodiment 1

[0135] Example 1. Preparation of C-peptide monoclonal antibody cross-linked magnetic particles

[0136] 1) Take 10 mg of 1.0 μm magnetic particles with active functional groups as carboxyl groups and wash them twice with 50 mM pH 6.0 MES buffer;

[0137] 2) After magnetic suction, remove the supernatant, add 0.5 mL of 50 mM pH 6.0 MES buffer and mix well, then add 0.5 mL of 25 mg / mL carbodiimide (EDC) solution and mix well;

[0138] 3) Reaction at room temperature for 30 minutes;

[0139] 4) Remove the supernatant after magnetic suction, and wash twice with 50mM pH 6.0 MES buffer;

[0140] 5) Add the 0.05 mg C peptide monoclonal antibody obtained in Example 1, and then dilute to 1 mL with 50 mM pH 6.0 MES buffer, and mix thoroughly;

[0141] 6) React overnight at 37°C;

[0142] 7) Wash twice with a solution containing 50mM pH 7.8Tris-HCl, 150mM NaCl, 1% BSA.

[0143] 8) Add 2 mL of a solution containing 50 mM pH 7.8 Tris-HCl, 150 mM NaCl, 1% BSA, and mix well.

[0144] 9) Block overnight at...

Embodiment 2

[0148] Example 2. Pretreatment of C peptide magnetic particle cross-linked product

[0149] 1) The non-ionic surfactant PE6400 and PE6200, diluted with purified water to 0.1% (mass percentage concentration)

[0150] 2) Example 1.6) After the procedure, magnetically remove the supernatant.

[0151] 3) Add C-peptide magnetic particle cross-linked product to 0.1% PE6400 solution or 0.1% In the PE6200 solution, the final concentration of the cross-linked magnetic particles is 10mg / mL; or add the cross-linked C-peptide magnetic particles to 0.1% PE6400 solution and 0.1% In the mixed solution of PE6200 solution (volume ratio 1:1), the final concentration of the cross-linked magnetic particles is 10 mg / mL, and mix for 90 minutes at 15-30°C;

[0152] 4) Continue the steps in Example 1.7) and beyond.

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
particle diameteraaaaaaaaaa
Login to View More

Abstract

The invention discloses a C-peptide monoclonal antibody cross-linked with magnetic particles, a preparation method thereof and a C-peptide test kit including the same. The C-peptide monoclonal antibody is pretreated with a non-ionic surfactant after cross-linked with the magnetic particles. The invention further discloses a C-peptide enzyme conjugate diluent with trehalose for improving the stability of the enzyme conjugate. The C-peptide test kit has the advantages of high sensitivity, good specificity, low drug interfere and good stability in results, thereby being applicable to clinical diagnosis and screening of diabetes widely.

Description

Technical field [0001] The invention relates to the field of in vitro diagnostic medical testing. Specifically, the present invention relates to a C-peptide monoclonal antibody cross-linked magnetic particle, a preparation method thereof, and a C-peptide detection kit including the same. Background technique [0002] C-peptide is a part of proinsulin synthesized by β islet cells in the pancreas. Before being secreted from islet cells, proinsulin mostly exists in the form of a polypeptide of 86 amino acids, including a polypeptide linked by two cystines, one is The 51 amino acid molecule of insulin is a 21 amino acid A chain and a 30 amino acid B chain, the other is a 30 amino acid molecule C peptide and two outside two 2 peptides. [0003] C peptide and insulin are secreted in equal amounts, but the concentration of C peptide in the blood is much higher than that of insulin. The main reasons are as follows: (1) C-peptide is different from insulin. It is not metabolized by the liv...

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): G01N33/68G01N33/577G01N33/553
CPCG01N33/531G01N33/553G01N33/577G01N33/68
Inventor 陈超李基彭波孙小禁
Owner SHANGHAI KEHUA BIO ENG
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