Expression Cloning of Antibodies from Single Human B Cells

Area of Science:

• Immunology
• Oncology
• Molecular Biology

Background:

• Most lymphomas arise from B cells, with antigen selection potentially driving their development.
• While B-cell receptors (BCRs) in chronic lymphocytic leukemia (CLL) show self-reactivity and proliferation, their role in other lymphomas is less understood.

Purpose of the Study:

• To develop and describe a strategy for characterizing the antibody reactivity of human B cells.
• To enable unbiased, single-cell level analysis of the human antibody repertoire.

Main Methods:

• Flow cytometric isolation of single human B cells.
• RT-PCR amplification of immunoglobulin (Ig) transcripts (IGH, IGK, IGL).
• Cloning of Ig transcripts into expression vectors for recombinant monoclonal antibody production.

Main Results:

• A detailed protocol for generating recombinant monoclonal antibodies from primary human B cells.
• The method allows for unbiased characterization of the antibody repertoire at the single-cell level.

Conclusions:

• This strategy facilitates the study of antibody reactivity in human lymphoma B cells.
• It provides insights into clonal evolution of B-cell lymphomas through single-cell sequencing of Ig transcripts.