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Hybridoma Technology

Hybridoma Technology

Hybridoma technology is used for the large-scale production of monoclonal antibodies. Monoclonal antibodies bind to only a single antigenic determinant or epitope. Such antibodies are used in research, diagnostics, and disease therapy. The hybridoma technology established in 1975 by Georges Köhler and Cesar Milstein was awarded the Nobel Prize in Medicine in 1984 for revolutionizing research and therapy.
Hybridoma Selection
Commonly used fusion techniques — electroporation,...

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Related Experiment Video

Updated: Apr 29, 2026

Antigenic Liposomes for Generation of Disease-specific Antibodies

Antigenic Liposomes for Generation of Disease-specific Antibodies

Published on: October 25, 2018

Immunization method for multi-pass membrane proteins using highly metastatic cell lines.

Hiroyuki Satofuka¹, Yoko Okabe¹, Yuki Takano²

¹Research and Development Division, Order-made Medical Research Inc., A-18 Green House, 2639 Yamazaki, Noda, Chiba 278-0022, Japan.

Biochemical and Biophysical Research Communications

|May 29, 2014

Summary

This summary is machine-generated.

Researchers developed a new method to create monoclonal antibodies (mAbs) targeting multi-pass membrane proteins using metastatic breast cancer cells. This approach successfully generated mAbs against the extracellular domain of solute carrier family 6 member 6 (SLC6A6).

Keywords:

Breast cancer Immunization method Metastasis Mouse monoclonal antibody

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Area of Science:

Immunology
Oncology
Biotechnology

Background:

Producing monoclonal antibodies (mAbs) against multi-pass membrane proteins is challenging due to their complex structure and difficulty in maintaining antigen integrity.
Existing methods face limitations in generating antibodies that target the extracellular domains of these proteins effectively.

Purpose of the Study:

To establish a novel method for producing monoclonal antibodies (mAbs) against multi-span membrane proteins using metastatic cancer cell lines.
To generate and characterize mAbs targeting the extracellular domain of solute carrier family 6 member 6 (SLC6A6), a 12 transmembrane protein, as a model.

Main Methods:

Grafting of metastatic breast cancer cell lines (MCF7-14 and MDA-MB231) into immunocompromised mice to induce immune responses.
Utilizing highly metastatic MDA-MB231 cells overexpressing SLC6A6 for immunization.
Screening and characterization of monoclonal antibodies using immunocytochemistry and flow cytometry.

Main Results:

Grafting metastatic cells induced significant splenic hypertrophy in mice.
Successfully produced seven IgG subclass mAb-producing clones that recognize the native extracellular domain of SLC6A6 on the cell surface.
The developed mAbs were confirmed to bind the extracellular domain of SLC6A6 in its native conformation.

Conclusions:

A novel immunization strategy using highly metastatic cells is effective for generating mAbs against multi-pass membrane proteins.
This method facilitates the development of therapeutic mAbs targeting challenging membrane protein antigens.
The approach holds promise for antibody development against various multi-pass membrane proteins relevant to disease.