CCP (Cyclic Citrullinated Peptide) used for detecting the immune antibody of rheumatoid arthritis with dot immuno-gold filtration assay

An immunodiafiltration and immune antibody technology, applied in the field of CCP peptides for the detection of rheumatoid arthritis antibodies, can solve problems such as waiting for further independent clinical research, no data to support BiP rheumatoid arthritis, and the impact of detection sensitivity.

Active Publication Date: 2013-06-19
SHANGHAI RONGSHENG BIOLOGICAL PHARM CO LTD
View PDF5 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

64% of patients with rheumatoid arthritis can produce antibodies directly against BiP, and there are reports that it is highly specific to the disease, but there is no data to support the role of BiP in predicting rheumatoid arthritis, and these reports The BiP antibody still needs to be confirmed by independent clinical research (Clinica Chimica Acta 2004, 350, 17-3

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
  • CCP (Cyclic Citrullinated Peptide) used for detecting the immune antibody of rheumatoid arthritis with dot immuno-gold filtration assay
  • CCP (Cyclic Citrullinated Peptide) used for detecting the immune antibody of rheumatoid arthritis with dot immuno-gold filtration assay
  • CCP (Cyclic Citrullinated Peptide) used for detecting the immune antibody of rheumatoid arthritis with dot immuno-gold filtration assay

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0085] The synthesis of embodiment 1 CCP peptide I, II and III

[0086] CCP peptides can be synthesized by solid-phase synthesis techniques using Fmoc chemistry. For the specific steps of this method, see Eur.J.Immunol.1994, 24, 3188-3193; J.Org.Chem.1972, 37, 3404-3409; Huang Weide, Chen Changqing Polypeptide Synthesis, Beijing: Science Press, 1985.

[0087] The formation method and steps of disulfide bonds can be found in the literature: Huang Weide, Chen Changqing Peptide Synthesis, Beijing: Science Press, 1985, p85; Michael W.Pennington Peptide Synthesis Protocols (Methods in Molecular Biology), Humana Press, 1994, p91-169 .

[0088] In this example, various CCP peptides were synthesized by Jill Biochemical (Shanghai) Co., Ltd., and the specific sequences are as follows:

[0089] CCP peptide I: His-Gln-Cys-His-Gln-Phe-Arg-Phe-Cit-Gly-Cit-Ser-Arg-Ala-Ala-Cys-Gly, on which the third and sixteenth Cys pass The sulfhydryl group forms a disulfide bond, and makes the polypept...

Embodiment 2

[0092] Example 2 Combination of CCP Peptides I, II and III with High Molecular Polymers

[0093] Dissolve 2 mg of carrier protein BSA in 200 μl of deionized water, and 2 mg of CCP peptides I, II, and III (molar ratio 1:1:1) in 0.5 ml of coupling buffer (0.1 mol / L MES, 0.9 mol / L NaCl, 0.2g / L NaN 3 , pH4.7). Add 0.5 ml of CCP solution to 200 μl of carrier protein solution. Add 1ml of 1-ethyl-(3-dimethylaminopropyl)carbodiimide (EDC) solution with a concentration of 10mg / ml into the mixture of CCP and BSA, mix gently, and react at room temperature for 2h , dialyzed against 0.01mol / L pH7.4 phosphate buffer at 4°C for 3 days to obtain the CCP peptide composition-BSA, which was stored at 4°C after aliquoting.

Embodiment 3

[0094] Example 3 IgG labeled colloidal gold

[0095] Heat 99ml ultrapure water to 80-90°C, add 1% (w / v) HAuCl 4 1ml, continue heating and stirring until boiling. Fill and boil for about 10 seconds, quickly add the existing trisodium citrate solution (0.0236g trisodium citrate, made into a concentration of about 1% (w / v)), stir and boil for 10 minutes. After cooling at room temperature, add ultrapure water to make up to 100ml. Scan the obtained colloidal gold at a wavelength of 400-700nm, and require the absorption value at a wavelength of 525-535nm to reach 0.92-0.98.

[0096] In 100ml of prepared gold sol that has been cooled to room temperature, use 1% (w / v) K 2 CO 3 Adjust the pH to 8.2.

[0097] Add 1.5 mg mouse anti-human IgG (Hangzhou Keelong Biotechnology Co., Ltd.) (0.1 M phosphate buffer diluted to 1 mg / ml), and stir at room temperature for 30 min.

[0098]Add BSA solution to block for 30min (5% (w / v) solution, add 25ml). Centrifuge at 10,000rpm for 25min and ...

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 relates to a CCP (Cyclic Citrullinated Peptide) used for detecting the immune antibody of rheumatoid arthritis with dot immuno-gold filtration assay, which comprises a CCP compound, a macromoleclar polymer, a millipore filter, a water absorption medium and a carrier medium, wherein the macromoleclar polymer is connected with the millipore filter in a covalent or noncovalent mode, and the CCP compound comprises CCPI, CCPII and CCPIII and is connected with the macromoleclar polymer in a covalent or noncovalent mode. The compound not only can realize the quick detection of the immune antibody of rheumatoid arthritis in vitro, but also can make the self-service diagnosis of patients easy, and the component also provides a reference to the timely understanding of the development of illness states.

Description

technical field [0001] The invention relates to a CCP peptide for detecting rheumatoid arthritis antibodies, in particular to a CCP peptide for detecting rheumatoid arthritis immune antibodies by a spot gold immunofiltration method, so as to realize rapid and accurate detection of rheumatoid arthritis immune antibodies. Background technique [0002] Clinically, autoimmune diseases mainly include systemic lupus erythematosus, rheumatoid arthritis, Sjogren's syndrome, dermatomyositis, polymyositis, systemic (multiple) sclerosis, scleroderma, etc. These diseases were once named " Connective tissue disease", later both abroad and domestically classified them as rheumatic diseases. [0003] Rheumatoid arthritis (RA) is a relatively common systemic autoimmune disease mainly manifested by chronic polyarticular inflammation. Very painful. The incidence of the disease is 0.5-1% worldwide, and the incidence in China is about 0.36%. The age of onset of patients is mostly between 20 ...

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
IPC IPC(8): C07K7/08G01N33/68G01N33/564
Inventor 朱绍荣
Owner SHANGHAI RONGSHENG BIOLOGICAL PHARM CO LTD
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