Cell fusion method for targeted capturing of CD138 single antibody

A technology of cell fusion and targeted capture, which is applied in the field of biomedicine and can solve problems such as cytotoxicity

Inactive Publication Date: 2018-07-24
翁炳焕
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Such an effective fusion dose of PEG is still highly toxic to cells

Method used

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  • Cell fusion method for targeted capturing of CD138 single antibody
  • Cell fusion method for targeted capturing of CD138 single antibody
  • Cell fusion method for targeted capturing of CD138 single antibody

Examples

Experimental program
Comparison scheme
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Embodiment Construction

[0012] figure 1 It is a schematic diagram of IgG fusion aiding of the present invention.

[0013] figure 2 is a cell fusion diagram of the present invention.

[0014] exist figure 1 Among them, 1 is myeloma cells expressing CD138, 2 is CD138 monoclonal antibody (IgG), and 3 is B lymphocytes expressing IgGFc receptor. The IgG Ab end of the CD138 monoclonal antibody binds to the CD138 receptor of myeloma cells, while the IgG Fc end binds to the B lymphocytes. Fusion under action.

[0015] exist figure 2 In this method, the fused cells were screened by HAT for 2 weeks, and photographed under an inverted microscope (40X) to obtain hybridoma cell clones.

[0016] Combine below figure 1 and figure 2 , the embodiments of the present invention are described in detail.

[0017] 1. Experimental reagents

[0018] (1) Pre-fusion agent: add 30ng / ml IL-3, 45ng / ml IL-6, 45ng / ml SCF, 3.6μg / L IFN-γ in DMEM medium, which is equivalent to the concentration of fused myeloma cells CD...

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Abstract

The invention provides a cell fusion method for targeted capturing of a CD138 single antibody and belongs to the field of biomedicines. The cell fusion method is characterized by comprising the following steps: putting a B cell to be fused and a myeloma cell into a pre-fusion agent which contains the CD138 single antibody capable of connecting the B cell and the myeloma cell, IL-3, IL-6 and SCF capable of activating the B cell, IFN-gamma capable of enhancing the activity of an IgGFc receptor of the B cell and concanavalin A which can be combined with a cell membrane receptor and is easily fused with a membrane, so that epitope CD138 of the myeloma cell is rightly combined with all the CD138 single antibody; the B cell is captured by an IgGFc fragment of the CD138 single antibody in a targeting manner to form a myeloma cell-CD138 single antibody-B cell conjugate, so that a cell distance is shortened and cell membrane contact and specific fusion are easy to realize; the specific fusion rate of the myeloma cell and the B cell is increased; invalid fusion between the B cell and the B cell, and between the myeloma cell and the myeloma cell is reduced.

Description

technical field [0001] The invention relates to a cell fusion method for CD138 monoclonal antibody target-specific capture in the field of biomedicine. Background technique [0002] Cell fusion is a cell engineering technology developed in the 20th century. It can make two or more cells (protoplasts) contact each other under certain conditions, and then undergo membrane fusion, cytoplasmic fusion and nuclear fusion to form hybrid cells. [0003] New cells formed by cell fusion (hybrid cells) have acquired genetic material from two parental cells and thus have new genetic or biological properties. Cell fusion has become one of the core technologies of cell engineering. It not only provides a powerful means for the study of nucleoplasmic relationship, gene regulation, genetic complementation, tumorigenesis, gene localization, aging control, etc. Plant distant hybrid breeding, developmental biology, immunology, medicine, food and agriculture and other fields have a wide range ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/06
CPCC12N15/02C12N2500/50C12N2501/125C12N2501/2303C12N2501/2306C12N2501/24C12N2501/599C12N2501/998
Inventor 翁炳焕李兰娟朱丹华朱小明温馨
Owner 翁炳焕
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