Unlock instant, AI-driven research and patent intelligence for your innovation.

Method for producing b cell mass, and method for producing monoclonal antibody employing same

A monoclonal antibody, B cell technology, applied in biochemical equipment and methods, animal cells, vertebrate cells, etc., can solve the problem that the mechanism is not fully elucidated, and achieve good efficiency.

Active Publication Date: 2019-11-05
KANEKA CORP +1
View PDF5 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

B cells that produce antibodies that display high affinity for antigens are known to be selected in the germinal center, but the mechanism of this selection has not been fully elucidated

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
  • Method for producing b cell mass, and method for producing monoclonal antibody employing same
  • Method for producing b cell mass, and method for producing monoclonal antibody employing same
  • Method for producing b cell mass, and method for producing monoclonal antibody employing same

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0038] The preparation method of the B cell population of the present invention includes: step c, while applying CD40 and BAFF receptor-mediated stimulation, in the absence of IL-21, in the absence of IL-4, and in addition to IL-21 and IL In the presence of cytokines other than -4, a cell population containing B cells is cultured together with a specific antigen; and, in step d, while Fas-mediated stimulation is applied, the cell population containing B cells is cultured to obtain a cell population containing cells that recognize the above-mentioned specific antigen. B cell population of B cells. The B cells recognizing a specific antigen prepared by the method of the present invention are preferably B cells specific for the specific antigen.

[0039]

[0040] Step c in the present invention is a step of applying stimulation mediated by CD40 and BAFF receptors in the absence of IL-21, in the absence of IL-4, and in the absence of IL-21 and IL-4. Cell populations comprising ...

Embodiment

[0117] (1) Cell culture

[0118] When there is no special statement, use RPMI-1640 (add 10% (v / v) FCS (fetal calf serum), penicillin / streptomycin, 2mM L-glutamine, 55nM 2-mercaptoethanol, 10mM HEPES and 1mM sodium pyruvate) as B cell culture medium in 5% (v / v) CO 2 , B cells were cultured at 37°C.

[0119] When there is no special statement, use D-MEM (add 10% (v / v) FCS, penicillin / streptomycin) as 40LB cell culture medium, in 5% (v / v) CO 2 The feeder cells and 40LB cells used for culturing B cells, which present CD40L and BAFF on the cell surface, were cultured at 37°C.

[0120] When conducting B cell culture experiments, 2.7×10 6 40 LB cells were inoculated in a 90 mm diameter cell culture plate (manufactured by Sumitomo Bakelite Co., Ltd.), cultured for 24 hours to form a monolayer, and then used after irradiating 120 Gy of X-rays.

[0121] (2) Preparation of feeder cells

[0122] FAIS (Fas-mediated Antigen-specific iGB-cell Selection) method for selecting antigen-spec...

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 present invention addresses the problem of providing a method for highly efficiently producing a B cell mass containing B cells capable of recognizing a specific antigen. According to the presentinvention, a method for producing a B cell mass is provided, which comprises: step c of culturing a cell mass containing B cells together with a specific antigen in the absence of IL-21, in the absence of IL-4 and in the presence of a cytokine other than IL-21 or IL-4 while applying a stimulus through CD40 and a BAFF receptor and step d of culturing the cell mass containing the B cells while applying a stimulus through Fas to produce a B cell mass containing B cells capable of recognizing the specific antigen.

Description

technical field [0001] The present invention relates to a method for producing a B cell population and a method for producing a monoclonal antibody using the same. Background technique [0002] Monoclonal antibodies showing high selectivity for specific antigens are expected to be developed into antibody drugs, and development of antibody drugs targeting cancer cells in particular is underway. In order to apply a drug containing a monoclonal antibody as an active ingredient to human therapy, from the viewpoint of avoiding rejection, it is most desirable to administer a human antibody with a small amount of xenoantigen. Therefore, various chimeric antibodies and humanized antibodies have been developed. [0003] After multiple times of immunizing mice with antigens, cells from the spleen and lymph nodes are fused with myeloma cells to form hybridomas, and based on the mouse IgG antibodies produced by the hybridomas, human therapeutic drugs are produced using recombinant tech...

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): C12N5/0781C12P21/08
CPCC07K16/00C07K16/24C07K16/32C07K2317/14C12N5/0635C12N2501/52A61K39/0011C07K16/2875C07K16/2878C12N2501/2302C12N2501/2304C12N2501/2312C12N2501/232C12N2501/2321C12N2501/2324C12N2501/2327C12N2501/25
Inventor 马渡达也中石智之北宽士北野光昭北村大介
Owner KANEKA CORP