Hybridoma Technology Explained: The Backbone of Monoclonal Antibody Production
Hybridoma technology is a cornerstone of modern biomedical research and diagnostic development. It enables the large-scale production of monoclonal antibodies—highly specific proteins that recognize a single epitope on a target antigen. Whether you’re working in basic research, therapeutic development, or in vitro diagnostics, understanding how hybridomas work can inform better decisions around antibody sourcing and project design.
At Green Mountain Antibodies, we specialize in hybridoma antibody production for research, diagnostics, and commercial applications. This article explains how the process works, highlights its advantages, and provides in-page links to relevant services and scientific references.
What Is Hybridoma Technology?
Hybridoma technology involves fusing antibody-producing B-cells from an immunized mouse with immortal myeloma cells. This fusion creates hybrid cells, or “hybridomas,” that inherit the specificity of the B-cell and the longevity of the cancerous myeloma cell. The technique was first developed by Köhler and Milstein in 1975 (Nature, 1975), earning them the Nobel Prize in 1984.
How Hybridoma Antibody Production Works
The process of hybridoma production includes several key steps:
- Immunization – A mouse is immunized with an antigen of interest over several weeks to generate a strong immune response.
- Cell Fusion – B-cells from the spleen are fused with immortal myeloma cells using polyethylene glycol (PEG) or electrofusion.
- Selection and Screening – Hybrid cells are cultured in selective media (e.g., HA medium), and supernatants are screened for antigen-specific antibodies.
- Cloning – Positive hybridomas are cloned by limiting dilution or fluorescence-activated cell sorting (FACS) to ensure monoclonality.
- Expansion and Cryopreservation – Monoclonal-producing hybridomas are expanded and frozen for long-term production.
This approach produces a renewable, consistent supply of monoclonal antibodies with high specificity and lot-to-lot consistency.
Why Hybridoma Technology Matters
Monoclonal antibodies derived via hybridoma are ideal for:
- Diagnostic assays like ELISA and lateral flow tests, where batch consistency is essential.
- Target validation in drug discovery pipelines.
- Basic research requires high-affinity, epitope-specific reagents.
Compared to polyclonal antibodies, hybridoma-derived monoclonals provide superior specificity and scalability.
References
Köhler G, Milstein C. Continuous cultures of fused cells secreting antibody of predefined specificity. Nature. 1975;256(5517):495–497. Link
Nelson AL, Dhimolea E, Reichert JM. Development trends for human monoclonal antibody therapeutics. Nat Rev Drug Discov. 2010;9(10):767–774. Link
Our Hybridoma Antibody Production Services
Green Mountain Antibodies offers custom hybridoma development services tailored to your research or diagnostic goals. Our U.S.-based lab supports:
- Immunogen design and peptide conjugation
- Hybridoma generation and screening
- Isotyping and large-scale production and sequencing
- Antibody purification and validation
We work closely with biotech and pharmaceutical teams to deliver high-affinity monoclonals with rapid turnaround and transparent project management.
Explore Our Hybridoma Development Services
Hybridoma technology remains a reliable, scalable, and reproducible method for monoclonal antibody generation. If you need consistent, high-specificity reagents for research or product development, our team is ready to help. Explore our hybridoma development services or contact us to discuss your next project.