Hybrid tumor, antihuman bladder cancer monoantibody and guide medicament, and bladder cancer diagnosis agent

A technology for bladder cancer and hybridoma, applied in the field of medicine and biology, can solve the problems of low sensitivity and specificity, achieve the effects of good specificity, avoid the production of anti-ricin A chain antibodies, and prevent recurrence

Inactive Publication Date: 2009-01-14
李翀
View PDF0 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Molecular markers newly used in clinical practice in recent years, such as nuclear matrix protein, bladder tumor antigen and satellite instability, are also limited by low sensitivity and specificity

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
  • Hybrid tumor, antihuman bladder cancer monoantibody and guide medicament, and bladder cancer diagnosis agent
  • Hybrid tumor, antihuman bladder cancer monoantibody and guide medicament, and bladder cancer diagnosis agent
  • Hybrid tumor, antihuman bladder cancer monoantibody and guide medicament, and bladder cancer diagnosis agent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Embodiment 1: Preparation of hybridoma cell lines

[0037] Material:

[0038] Human bladder cancer cell line E-J cells (Beijing Northern Weiye Development Co., Ltd.)

[0039] Balb / C mice (Beijing Weitong Lihua Experimental Animal Technology Co., Ltd.)

[0040] Mouse myeloma cells Sp2 / 0 (ATCC CRL-1772)

[0041] method:

[0042] 1) Immunization: Immunize Balb / C mice with human bladder cancer cell line E-J cells at 1×10 7cells / 0.5ml PBS for intraperitoneal injection. Mice were re-immunized 2 weeks later with the same injection volume and method. After the mouse serum titer reaches the requirement, the cell fusion is prepared, and the mice are boosted three days before the fusion. Mouse myeloma cells Sp2 / 0 (ATCC CRL-1772) were prepared at the same time as the mice were immunized.

[0043] 2) Cell fusion: The sensitized B lymphocytes and myeloma cells were fused according to the following method:

[0044] (1) Take logarithmically grown myeloma cells Sp2 / 0, centrifuge...

Embodiment 2

[0054] Embodiment 2: Preparation and purification of DTB monoclonal antibody

[0055] The above-mentioned hybridoma cell DTB was inoculated into the peritoneal cavity of mice to prepare ascites, and monoclonal antibodies were extracted from the ascites. Purification of monoclonal antibody DTB: using Protein G affinity chromatography. First prepare the Protein G affinity chromatography column, equilibrate the column with PBS, take the ascites fluid containing DTB monoclonal antibody to pass through the column, then wash the column with PBS until the OD value is close to zero, and use 0.2M glycine-HCl solution (pH2. 8) Elution, collect the eluate, measure the OD value of each collection tube, keep the eluate in the peak area, and use the eluate to prepare the immunoconjugate after being concentrated by dialysis.

Embodiment 3

[0056] Embodiment 3: Characterization of DTB monoclonal antibody

[0057] Immunohistochemical staining was performed on human bladder cancer tissue sections according to conventional methods, and the results were as follows: figure 1 , 2 shown. The results showed that the human bladder cancer tissue showed positive reaction after staining with DTB monoclonal antibody, secondary antibody and substrate, while the normal human bladder tissue showed negative reaction after staining with DTB monoclonal antibody, secondary antibody and substrate.

[0058] Immunofluorescence was performed on bladder cancer cells E-J and other cells using DTB monoclonal antibody according to conventional methods. The result is as figure 1 , 2 shown. The results showed that the DTB monoclonal antibody had a strong positive reaction with E-J cells, and had no cross-reaction with other non-bladder cancer cells.

[0059] Table 1 Reactivity of DTB monoclonal antibody with human cell lines

[0060] ...

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
Sensitivityaaaaaaaaaa
Login to view more

Abstract

The invention relates to a hybridoma which can generate a monoclonal antibody against human bladder cancer, and parental cells of which are B lymphocytes of a mouse and myeloma cells of the mouse immunized by E-J cells of a human bladder cancer cell line. The invention also relates to purposes and a preparation method of the hybridoma, the monoclonal antibody generated by the hybridoma, a directional drug against bladder cancer and a bladder cancer immunological diagnostic reagent and a preparation method of the directional drug. The antibody part of the directional drug serves as the monoclonal antibody against human bladder cancer. The diagnostic reagent is composed of an E-J cell lysis liquid of the human bladder cancer cell line and the monoclonal antibody DTB against human bladder cancer coated on an enzyme label plate, and the bladder cancer antigen in urine exfoliated cells is examined through adopting the competitive ELISA method. The specificity of the antibody of the hybridoma is strong; the tumor killing function of the directional drug of the hybridoma is strong without drug tolerance but good specificity; and the diagnostic reagent of the hybridoma has the advantages of damage resistance, high sensibility and specificity as well as convenience and quickness.

Description

technical field [0001] The present invention relates to the field of medical biotechnology, in particular to a hybridoma DTB producing anti-human bladder cancer monoclonal antibody, its use and preparation method, anti-human bladder cancer monoclonal antibody DTB, guiding drugs and bladder cancer immunodiagnostic reagents, And the preparation method of the targeting drug. Background technique [0002] Bladder cancer is the most common tumor in the urinary system. Although multiple measures are currently used for comprehensive treatment, 40%-70% of patients still have one or more recurrences, and 10%-15% of patients develop higher-grade tumors or metastases. There are also obvious toxic side effects. [0003] Prevention of recurrence of bladder cancer after tumor resection is an important clinical issue. The medicines currently used for intravesical perfusion to prevent recurrence are mainly divided into the following categories: (1) antitumor chemotherapeutic drugs, such ...

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): C12N5/20C07K16/18A61K39/395A61K38/16A61P35/00G01N33/577
Inventor 李翀侯强王德朋栗刚
Owner 李翀
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