Check patentability & draft patents in minutes with Patsnap Eureka AI!

A kind of tumor stem cell detection kit

A kit and a specific technology, applied in the field of tumor stem cell detection kits, can solve the problems of low early warning signals, high missed diagnosis rate, and many complications in early detection

Active Publication Date: 2020-08-07
河北意和医学检验实验室有限公司
View PDF1 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the limitations of examination at the cellular level, such as fecal occult blood test (FOBT) and invasive colonoscopy combined with pathological biopsy, the early detection and early warning signals are low, and the missed diagnosis rate is high, and there are many complications. factors; thus forcing researchers to find effective non-invasive methods from the molecular level[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
  • A kind of tumor stem cell detection kit
  • A kind of tumor stem cell detection kit
  • A kind of tumor stem cell detection kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1. Preparation of DCLK1 protein antigen

[0034] The difference between the short isomer of DCLK1 (SEQ ID NO:1) and the long isomer (SEQ ID NO:2) is only that the first 6 amino acids of the short isomer are different from those of the long isomer. Therefore, we used the first 9 amino acids of the short isomer (SEQ ID NO: 3) as an antigen to prepare an antibody that only specifically binds to the short isomer of DCLK1. Artificially synthesized a plasmid containing 6 repeats of SEQ ID NO:3 (purchased from Beijing Qingke Biotechnology Co., Ltd.), denoted as pET-43.1b(+)-sDCLK1-6, the synthesis technology is known in the art. Transform the above plasmids into Escherichia coli competent BL21 (DE3), then add the competent cells to 2ml LB culture containing ampicillin resistance for overnight culture, and inoculate 1% inoculum in 20 mL LB containing ampicillin resistance on the next day In LB medium, shake culture at 37°C for 3 h, then add IPTG with a final concentrat...

Embodiment 2

[0035] Example 2. Preparation and purification of anti-DCLK1 antibody

[0036] The antigen prepared in Example 1 was injected intraperitoneally into female Balb / c mice together with Freund's complete adjuvant that caused an immune response for initial immunization, and then the same amount of antigen was injected into the above-mentioned female Balb / c mice every 2 weeks Mice were immunized intraperitoneally. Finally, the final immunization was carried out 3 days before the fusion with myeloma cells. After mixing 100 μg of the DCLK1 antigen in Example 1 with an immune adjuvant, the mice were given intraperitoneal final immunization. The immunized Balb / c mouse splenocytes were taken and fused with the myeloma Sp2 / 0 cell line, and the fused cells were treated with Iscove medium (0.1 mM hypoxanthine, 0.4 μM aminopterin) containing 10% serum and 16 μM thymidine), diluted to an appropriate concentration, added to a 96-well plate for cultivation. After 10 days, the cell supernatant...

Embodiment 3

[0037] Example 3. Anti-DCLK1 antibody subtype identification

[0038] The mouse monoclonal antibody subtype identification kit (Proteintech) was used to identify the subtype of the anti-DCLK1 monoclonal antibody obtained in Example 2. The ELISA plate provided in the kit is pre-coated with specific antibodies against mouse IgG1, IgG2a, IgG2b, IgG2c, IgG3, IgM, kappa light chain, lambda light chain, and the anti-DCLK1 antibody purified in Example 2 Samples were added to the sample wells, 50 μl per well. Without incubation, add 1X goat anti-mouse IgM+IgG-HRP to 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, develop color for 15 minutes at room temperature in the dark, and add 100 μl stop solution. Detect the OD value at 450nm by microplate reader, the result is as follows figure 1 As shown, the hea...

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 tumor stem cell detection kit, which comprises an antibody capable of specifically binding to short isoform DCLK1 protein, can indicate the existence and distribution of colon cancer stem cells, and has broad application prospects.

Description

technical field [0001] The invention relates to the field of biological detection, in particular to a tumor stem cell detection kit. Background technique [0002] In China, colorectal cancer is the fourth most common cancer and the fifth most common cause of death, with an estimated 331,300 new cases and 159,300 deaths per year[1], and its incidence increases with the age of patients constant increase. With the deepening of research, it is increasingly recognized that most colorectal cancer (CRC) originates from colorectal adenoma (CRA), which is an abnormal growth and tumor on the surface of the colorectum. Protrusion is a common disease in gastroenterology. Adenomas develop slowly and sometimes undergo malignant transformation through a series of genetic and epigenetic alterations, and progression from detectable adenoma to cancer can take at least 10 years [2]. However, due to the insidious onset of the disease and the lack of specificity in clinical manifestations, it...

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): C07K16/40G01N33/577G01N33/574G01N33/573
CPCC07K16/40C07K2317/56C07K2317/565C07K2317/92G01N33/573G01N33/57419G01N33/57492G01N33/577G01N2333/91215
Inventor 王阳金鑫
Owner 河北意和医学检验实验室有限公司
Features
  • R&D
  • Intellectual Property
  • Life Sciences
  • Materials
  • Tech Scout
Why Patsnap Eureka
  • Unparalleled Data Quality
  • Higher Quality Content
  • 60% Fewer Hallucinations
Social media
Patsnap Eureka Blog
Learn More

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

© 2025 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com