Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Kit for rapidly detecting whether hybridoma cells secrete antibodies or not

A hybridoma cell and cell secretion technology, which is applied in the field of kits for rapid detection of whether hybridoma cells secrete antibodies, can solve the problems of unstable passage, lack of cross-linking function, and loss of antibody production.

Pending Publication Date: 2022-05-13
ZHEJIANG ZHENGXI BIOMEDICAL CO LTD
View PDF0 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, each hybridoma cell is the product of the fusion of B cells and myeloma cells, which is unstable during cell division and prone to lose the ability to produce antibodies
The absence of the Fc segment of the antibody results in a smaller F(ab) segment and lack of cross-linking function, so that it cannot bind to the antigen and undergo a precipitation reaction, nor will it be captured by immune cells in in vivo studies

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
  • Kit for rapidly detecting whether hybridoma cells secrete antibodies or not
  • Kit for rapidly detecting whether hybridoma cells secrete antibodies or not
  • Kit for rapidly detecting whether hybridoma cells secrete antibodies or not

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1 A method for rapidly detecting antibodies secreted by hybridoma cells

[0045] The cells to be detected are CD22[10F.4.4.1], and the type of antibody expressed: Mouse IgG.

[0046] Resuscitated cells, counted after rejuvenation, and took 4×10 6 Each cell to be tested was centrifuged at 1500rpm / min for 5min, then washed with 10mL PBS buffer, and centrifuged at 1500rpm / min for 5min. After centrifugation, 200 μL of RPMI-10% complete culture solution containing protein transport inhibitor was added.

[0047] RPMI-10% complete culture solution (500mL) formulation is shown in the following table:

[0048]

[0049]The protein transport inhibitor is a protein transport inhibitor containing brefedex A (555029 / BD Biosciences), and the formula is 6 mL of RPMI-10% complete culture solution, adding 600 μL of a protein transport inhibitor containing brefexedin A, resuspended cells by pipetting, Pipet into a round-bottom 96-well plate in a carbon dioxide incubator at 3...

Embodiment 2

[0056] Referring to the experimental method of Example 1, CD22[5E8.1.8] was detected, and CD22[5E8.1.8] expressed MouseIgG.

[0057] Cells stained with CD22[5E8.1.8] accounted for 93.976% of the total cells ( image 3 ).

Embodiment 3

[0059] Referring to the experimental method of Example 1, CD3[SK7] was detected, and CD3[SK7] expressed Mouse IgG1, kappa.

[0060] CD3[SK7] cells accounted for 0.352% of the total cells ( Figure 4 ).

[0061] Judging from the above examples, CD22[5E8.1.8] cells had the highest positive rate of secreting antibodies, followed by CD22[10F4.4.1], and CD3[SK7] cells had no positive cells secreting antibodies.

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 method for rapidly detecting the antibody secreted by the hybridoma cell comprises the following steps: displaying an Fc segment of the antibody in the hybridoma cell in cytoplasm, analyzing the Fc segment in the cytoplasm, and judging the antibody specificity and the antibody generation capability of the cell. According to the method provided by the invention, the positive rate proportion of antibody cells secreting antibodies in some intracellular expression antibodies and hybridoma cells in the total cell number can be visually detected. The antibody secretion capability of each monoclonal cell and the non-specific component on the surface of the hybridoma cell can be detected.

Description

technical field [0001] The invention belongs to the field of cell biology, in particular to a kit for rapidly detecting whether hybridoma cells secrete antibodies. Background technique [0002] An antibody is an immunoglobulin (Ig) that specifically binds an antigen, which can be hydrolyzed by proteolytic enzymes. For example, it is hydrolyzed into two F(ab) segments and one Fc segment by papain. The F(ab) segment of the antibody can bind to the antigen, while the Fc segment functions to bind to the Fc receptor and mediate opsonization or antibody-dependent cell-mediated cytotoxicity (ADCC, antibody-dependent cell-mediated cytotoxicity ). Since the Fc segment can be directly combined with enzymes or fluorescent dyes to label antibodies, it is not only the site of antibody rivets on the fixed plate in ELISA experiments, but also the site of recognition and binding of secondary antibodies in immunoprecipitation, western blotting and immunohistochemistry. [0003] Hybridoma ...

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): G01N33/68
CPCG01N33/6854G01N33/56966
Inventor 张洋褚建达张娅
Owner ZHEJIANG ZHENGXI BIOMEDICAL CO LTD
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com