Antigen-specific human antibodies from mice comprising four distinct genetic modifications

Area of Science:

• Immunology
• Biotechnology
• Genetic Engineering

Background:

• Human monoclonal antibodies offer therapeutic potential due to high specificity and low immunogenicity.
• Current methods struggle to reliably produce high-affinity human antibodies, necessitating the use of rodent-derived antibodies.
• Rodent antibody systems have advantages like ease of immunization and lack of tolerance to human antigens.

Purpose of the Study:

• To develop a reliable method for generating high-affinity human antibodies.
• To overcome the limitations associated with using rodent antibodies in clinical settings.
• To create transgenic mice capable of producing a diverse repertoire of human antibodies.

Main Methods:

• Development of transgenic mice lacking endogenous mouse immunoglobulin expression (IgM, Ig kappa).
• Introduction of human immunoglobulin transgenes (IgM, IgG, Ig kappa) into mice.
• Disruption of endogenous V(D)J recombination in heavy and kappa light chain loci.
• Immunization of transgenic mice with human proteins and isolation of hybridomas.

Main Results:

• Transgenic mice successfully generated human sequence immunoglobulins through V(D)J joining, class switching, and somatic mutation.
• Homozygous disruption of endogenous loci ensured exclusive production of human antibodies.
• Hybridomas secreting human IgG kappa antigen-specific antibodies were successfully isolated after immunization.

Conclusions:

• The developed transgenic mice provide a robust platform for producing human sequence monoclonal antibodies.
• This technology addresses the need for high-affinity human antibodies, advancing therapeutic antibody development.
• The system combines the advantages of mouse immunization with the production of fully human antibodies.