Colorectal cancer detection kit

A kit and quantitative detection technology, applied in the field of immunoassay, to achieve the effect of simple microplate reader, fast and sensitive detection, simple and easy operation

Active Publication Date: 2021-06-01
广东明志医学检验实验室有限公司
View PDF5 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is still a need for monoclonal antibodies with higher affinity and more favorable properties

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
  • Colorectal cancer detection kit
  • Colorectal cancer detection kit
  • Colorectal cancer detection kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1. Preparation and purification of anti-CEA antibody

[0023] CHO cells stably expressing CEA were cultured with DMEM medium, and 10 7 CHO-CEA + The cells were injected intraperitoneally into female Balb / c mice together with Freund's complete adjuvant for initial immunization, followed by 10 injections every 2 weeks 7 CHO-CEA + The cells were injected intraperitoneally into the above-mentioned female Balb / c mice, and the immunization was continued. Finally, the last immunization was carried out 3 days before the fusion with myeloma cells, and the final immunization was carried out with 10 7 CHO-CEA + Cells were immunized intravenously. Get immunized Balb / c mouse splenocytes, fuse it with myeloma Sp2 / 0 cell line, and use Iscove medium (0.1 mM hypoxanthine, 0.4 μ M aminopterin) to contain 10% serum after fusion and 16 μM thymidine), diluted to an appropriate concentration, added to a 96-well plate for cultivation. After 10 days, the cell supernatant was tak...

Embodiment 2

[0025] Example 2. Anti-CEA antibody subtype identification

[0026] The subtype of the anti-CEA monoclonal antibody Cab-207 obtained in Example 1 was identified by ELISA (the kit was purchased from Proteintech). The ELISA plate provided in the kit has been pre-coated with specific antibodies against mouse IgG1, IgG2a, IgG2b, IgG3, IgA, IgM, kappa light chain, lambda light chain, and the anti-CEA purified in Example 1 Antibody sample Cab-207 was added to the sample well, 50 μl per well, without incubation. Add 1X goat anti-mouse IgA+IgM+IgG-HRP into the sample wells, 50 μl per well, mix gently, and incubate for 1 h. Remove the liquid in the well and add 1XPBST to wash the well 3 times, and absorb excess water with absorbent paper. Add chromogenic solution, 100 μl per well, and develop color for 15 minutes at room temperature in the dark. Add 100 μl stop solution to stop the color reaction. Detect the OD value at 450nm by microplate reader, the result is as follows figure 1...

Embodiment 3

[0027] Example 3. Monoclonal Antibody Sequence Determination

[0028] After recovering the hybridoma cell line corresponding to Cab-207, culture it to a total number of 10 7 Cells were collected by centrifugation at 1000rpm for 5min, and RNA was extracted. Add TRNzol-A+ to the cell pellet for lysis, and let it stand at room temperature for 15 minutes. Add 200μl chloroform per ml TRNzol-A+, vortex for 15 seconds, and let stand for 3 minutes. Centrifuge at 13000rpm at 4°C for 10 minutes. Trizol-A+ cell solution is divided into three layers: the upper colorless aqueous phase, the middle layer and the lower yellow organic phase. Transfer the aqueous phase containing RNA to a centrifuge tube, and add an equal volume of isopropanol, mix well, and let stand at room temperature for 25 minutes. Centrifuge at 13,000 rpm at 4°C for 10 minutes, discard the waste liquid to obtain the RNA pellet that sinks to the bottom. After the RNA pellet was washed twice with 75% ethanol, the RNA wa...

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 the field of tumor immunodetection, in particular to a colorectal cancer detection kit, and further discloses an application of an antibody to preparation of a tumor treatment and / or diagnosis reagent. The application provides an effective tool for diagnosis of prognosis judgment, curative effect observation and monitoring of metastasis or relapse of tumor focuses for colon cancer and rectal cancer patients.

Description

technical field [0001] The invention relates to the technical field of immunodetection, in particular to a colorectal cancer detection kit. Background technique [0002] Colorectal cancer is one of the most common tumors in the world. In developed countries in Europe and the United States, its incidence rate ranks second among malignant tumors, and ranks third in the world, second only to lung cancer and gastric cancer in men, and second only to breast cancer and cervical cancer in women. Colorectal cancer is most common in North America, Western Europe, Australia and New Zealand. In my country, the incidence of colorectal cancer ranks fifth among men and fifth among women. Compared with developed countries, the age of onset of colorectal cancer in my country is significantly earlier, and the median age of onset is about 45 years old. Moreover, with the improvement of living standards in recent years, the incidence of colorectal cancer continues to maintain a strong upwar...

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): C07K16/30G01N33/574G01N33/577
CPCC07K16/3007C07K2317/52C07K2317/56C07K2317/565G01N33/57419G01N33/57473G01N33/577
Inventor 王阳金鑫
Owner 广东明志医学检验实验室有限公司
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