Monoclonal antibodies (mAbs) are powerful tools used in both research and clinical applications. Producing mAbs requires a complex process that involves many different steps, from the initial hybridoma cell production to the final purification.
In this article, you’ll be provided an overview of the entire monoclonal antibody production process for those who are new to this field.
Creating Hybridomas
The first step in creating mAbs is to create hybridoma cells that are capable of producing them. To do this, scientists must first immunize an animal with an antigen of interest, such as a cancer-associated protein or toxin, and then collect immune cells from the animal’s blood.
Then, these cells are fused together with myeloma cells to form a stable hybridoma cell line that can produce large amounts of antibodies against the antigen.
Cloning and Expansion
Once the hybridoma cell line is created, it needs to be propagated and cloned to create enough material for downstream analysis and manufacturing purposes. This is usually done through serial dilution culturing or limiting dilution culturing methods in order to ensure that each clone has the same characteristics as its parent cell line.
Purification and Characterization
Once enough material is produced, it needs to be purified and characterized before it can be used for research or clinical applications. During this stage of the monoclonal antibody production process, impurities such as other proteins or contaminants are removed from the sample so that only pure mAbs remain.
Once purified, the sample is characterized using techniques such as SDS-PAGE or Western blotting in order to determine its identity and purity levels.
