Generation of Recombinant Human IgG Monoclonal Antibodies from Immortalized Sorted B Cells

作者： Gisela Nogales-Gadea*1,2, Abhishek Saxena*1, Carolin Hoffmann*1, Judith Hounjet1, Daniëlle Coenen1, Peter Molenaar1, Mario Losen1, Pilar Martinez-Martinez1 1School for Mental Health and Neuroscience,Maastricht University, 2Department of Neurosciences,Institut d'Investigació Germans Trias i Pujol

简介：

Overview

This article presents a protocol for synthesizing human monoclonal antibodies from B cells. The process involves isolating B cells, selecting those producing IgG, and generating recombinant antibodies for research on immune responses.

Key Study Components

Area of Science

• Immunology
• Antibody Engineering
• Cell Biology

Background

• Monoclonal antibodies are crucial for studying immune mechanisms.
• Understanding auto-antibody-mediated responses can lead to better therapies.
• Isolation and immortalization of B cells are key steps in antibody production.
• Protocols for generating human IgG are essential for research applications.

Purpose of Study

• To develop a reliable method for producing human monoclonal antibodies.
• To facilitate research into the mechanisms of immune responses.
• To provide a framework for future studies on auto-antibodies.

Methods Used

• Isolation of B cells from blood or thymus tissue.
• Selection of IgG-positive B cells using flow cytometry.
• Immortalization of selected B cells via Epstein-Barr virus.
• RNA isolation and cDNA generation for antibody cloning.

Main Results

• Successful isolation and immortalization of B cells.
• Generation of high quantities of recombinant human IgG.
• Validation of IgG production through ELISA testing.
• Establishment of a protocol for future antibody studies.

Conclusions

• The developed protocol enables efficient production of human monoclonal antibodies.
• This method can advance research on auto-antibody-mediated immune responses.
• Future applications may include therapeutic development and diagnostic tools.

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