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Related Experiment Video

Updated: May 18, 2026

Characterization of Thymus-dependent and Thymus-independent Immunoglobulin Isotype Responses in Mice Using Enzyme-linked Immunosorbent Assay

Characterization of Thymus-dependent and Thymus-independent Immunoglobulin Isotype Responses in Mice Using Enzyme-linked Immunosorbent Assay

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A method for inducing antigen-specific IgG production by in vitro immunization.

Mieko Kato¹, Huimin Yan, Noriko M Tsuji

¹Molecular Composite Medicine Research Group, Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), 1-1-1 Higashi, Tsukuba, 305-8566, Japan.

Journal of Immunological Methods

|September 15, 2012

Summary

This study presents an improved in vitro immunization (IVI) protocol using mouse spleen cells to increase antigen-specific IgG antibody production. The enhanced IVI method boosts plasma cell frequency and antibody affinity for broader applications.

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Last Updated: May 18, 2026

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

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

Published on: June 5, 2015

Area of Science:

Immunology
Biotechnology

Background:

In vitro immunization (IVI) offers advantages over conventional methods but suffers from low positive clone numbers and limited IgG production.
Current IVI protocols often yield predominantly IgM antibodies, restricting their therapeutic and diagnostic applications.

Purpose of the Study:

To develop an optimized in vitro immunization protocol for enhanced generation of antigen-specific IgG antibodies from mouse spleen cells.
To improve the frequency and affinity of antibody-producing plasma cells generated through in vitro immunization.

Main Methods:

Employing multiple cycles of repeated antigen stimulation and subsequent cell expansion.
Optimizing culture conditions, including cell density, stimulant type, and initial cell preparation.

Main Results:

The improved IVI protocol significantly increased the frequency of plasma cells producing antigen-specific IgG antibodies.
Gene and cytokine expression analysis indicated antigen-specific B cell activation via CD4-positive helper T cells.
Successfully generated an IgG antibody against keyhole limpet hemocyanin with a dissociation constant of 10(-7)M.

Conclusions:

The optimized IVI protocol effectively overcomes limitations of previous methods, enabling efficient IgG antibody production.
This enhanced IVI approach provides a valuable tool for generating high-affinity IgG antibodies for research and development.