Nanometer magnetic particle chemiluminescence detection kit for carbohydrate antigen CA19-9 as well as preparation method thereof and detecting method thereof

A technology of nano-magnetic particles and chemiluminescence, which is applied in chemiluminescence/bioluminescence, analysis and measurement devices through chemical reactions of materials, and can solve problems such as poor process stability, limited detection effect, and cumbersome preparation process , to achieve cost advantages, simple and time-saving operation, and high sensitivity

Active Publication Date: 2014-11-26
SUZHOU HAOOUBO BIOPHARML
View PDF6 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the kit can realize rapid and accurate detection, the preparation process of the solid phase carrier coated with CA19-9 monoclonal antibody and the horseradish peroxidase-labeled CA19-9 monoclonal antibody is very cumbersome and the process The stability is not good, which limits its detection effect, especially the precision between analyzes and increases the cost

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
  • Nanometer magnetic particle chemiluminescence detection kit for carbohydrate antigen CA19-9 as well as preparation method thereof and detecting method thereof
  • Nanometer magnetic particle chemiluminescence detection kit for carbohydrate antigen CA19-9 as well as preparation method thereof and detecting method thereof
  • Nanometer magnetic particle chemiluminescence detection kit for carbohydrate antigen CA19-9 as well as preparation method thereof and detecting method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Example 1 Preparation of the first reagent

[0054] (1) Materials and equipment: Carbohydrate-related antigen CA19-9 (CA19-9) monoclonal antibody stored in phosphate buffer (purity over 95wt%, concentration 2mg / mL); fluorescein isothiocyanate (FITC) ), sodium carbonate and other reagents should be chemically pure; G-25 gel purification column is purchased from GE.

[0055] (2) Preparation steps:

[0056] ① Prepare 0.5mg / mL FITC solution with 0.1~0.2mol / L carbonate buffer with pH 9.0~10.0;

[0057] ②According to the ratio of CA19-9 antibody to FITC molecule ratio of 1:20, add the FITC solution prepared in step ① to the antibody solution, mix well, and let stand at room temperature for 12 hours to react to generate CA19-9 antibody-FITC conjugate;

[0058] ③ Separate the reaction solution after step ② through a G-25 gel column to remove unreacted FITC, and obtain a solution containing CA19-9 antibody-FITC conjugate (ie FITC-labeled CA19-9 antibody);

[0059] ④Dilute the solution con...

Embodiment 2

[0060] Example 2 Preparation of the second reagent

[0061] (1) Materials and equipment: CA19-9 monoclonal antibody stored in phosphate buffer (purity over 95wt%, concentration of 2mg / mL); alkaline phosphatase stored in phosphate buffer (ALP solution, ALP purity is About 99%, the specific activity is about 1500U / mg, the concentration is 10mg / mL); the cross-linking agent SMCC, 2-IT is purchased from THERMO, the chemical reagents such as TRIS should be chemically pure; the G-25 gel purification column is GE company's product.

[0062] (2) Preparation steps:

[0063] ①Take 1mg of CA19-9 antibody, add 2-4μL of 10mg / mL coupling agent 2-IT solution, leave it at room temperature for 20min, add 10μL of 0.1mol / L glycine solution, leave it at room temperature for 5min, use G-25 gel Desalt the column, collect the activated CA19-9 antibody, store it at 2-8°C for later use;

[0064] ②Take 1.5mg of ALP solution, add 10-20μL of 5mg / mL SMCC solution, let stand at room temperature for 30min, remove ...

Embodiment 3

[0067] Example 3 Preparation of magnetic separation reagent

[0068] (1) Materials and instruments:

[0069] Magnetic particle suspension: magnetic particle content 5wt%, magnetic particle contains carboxyl group (COOH) active group, carboxyl group content per gram (g) magnetic particle (dry weight) is not less than 0.4 millimoles (mmol), with super paramagnetism, The diameter is between 0.5-2μm.

[0070] Anti-FITC antibody: It can be a polyclonal antibody or a monoclonal antibody, with a purity of more than 90wt%, and a dilution titer of more than 1:1 million;

[0071] 2-Morpholine ethanesulfonic acid (MES), carbodiimide (EDC), TRIS and other reagents should be chemically pure.

[0072] (2) Preparation steps:

[0073] ① Take a suspension of 100 mg of magnetic particles, magnetically separate the supernatant, and resuspend in 10 mL of 0.05 mol / L, pH 4.5~5 MES buffer;

[0074] ②Add 2~4mg of anti-FITC antibody and suspend at room temperature for 30~60min;

[0075] ③Add 0.5~1mL of a freshly ...

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
diameteraaaaaaaaaa
Login to view more

Abstract

The invention relates to a nanometer magnetic particle chemiluminescence detection kit for a carbohydrate antigen CA19-9 as well as a preparation method thereof and a detecting method thereof. The kit comprises a solution, which contains a fluorescein labeled carbohydrate related antigen CA19-9, a suspension, which is coated with a fluorescein antibody, and a solution, which contains an alkaline phosphatase labeled carbohydrate related antigen CA19-9 antibody, wherein the alkaline phosphatase labeled carbohydrate related antigen CA19-9 antibody is obtained by connecting an alkaline phosphatase and the carbohydrate related antigen CA19-9 antibody through cross-linking agents SMCC and 2-IT. According to the invention, the carbohydrate related antigen CA19-9 can be quantitatively detected with lower cost, higher accuracy and higher precision.

Description

Technical field [0001] The invention belongs to the field of biotechnology, and specifically relates to a nano-magnetic particle chemiluminescence assay kit of carbohydrate-related antigen CA19-9 that combines immunomagnetic particle separation technology and chemiluminescence immunoassay technology, and a preparation method and detection method thereof. Background technique [0002] Carbohydrate Antigen 19-9 (Carbohydrate Antigen19-9) is a glycolipid on the cell membrane with a molecular weight of about 1000KD. CA19-9 was named by Koprowski et al. in 1979 by immunizing mice with colon cancer cells and hybridizing with myeloma to obtain the 116NS19-9 monoclonal antibody. It is the most sensitive marker for pancreatic cancer reported so far. [0003] CA19-9 is mainly used clinically in the following areas: [0004] Malignant tumors of the digestive tract [0005] The CA 19-9 levels of pancreatic cancer, hepatobiliary cancer, gastric cancer, and colorectal cancer were 683, 535, 279, an...

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): G01N33/574G01N21/76
Inventor 于大为程晓蕾赵文姬
Owner SUZHOU HAOOUBO BIOPHARML
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