A Guide to the Monoclonal Antibody Production Process
The production of monoclonal antibodies (mAbs) involves the use of cell cultures with optimized conditions for growth to generate a target molecule. Then, various additional steps follow to isolate and purify this molecule. At Cell Culture Company, we are here to support you in the monoclonal antibody development process.
Monoclonal antibodies may be produced in multiple ways, including through the use of hybridoma technology, in vitro synthetic genes (to produce recombinant mAbs), existing hybridoma (to produce recombinant mAbs), and phage display (to produce recombinant mAbs). Other methods include B cell cloning and Next-Generation Sequencing, which is not covered here.
Using Hybridoma Technology
Monoclonal antibody production usually begins with the immunization of an animal with the target antigen to bring about an immune response which involves the animal’s B cells generating antigen-specific antibodies. Once immunization has occurred antibody producing cells are harvested from the spleen and joined with tumor cells to develop hybridomas which are screened for the production and performance of antibodies.
Through isolation, the hybridoma cells producing antibodies are cloned and then cultivated with the application of cell culture methods. The cells secrete antibodies into the culture media. These antibodies may be harvested and purified via affinity purification or used in their crude form.
Using In Vitro Synthetic Genes (for Recombinant mAbs)
When compared to traditional polyclonal and monoclonal antibodies, recombinant antibodies provide a secure, long-term source of antibodies with negligible variation between batches. The production of recombinant antibodies is carried out through the cloning of antibody-coding genes into a mammalian expression vector. Functional antibodies are manufactured as these vectors are transferred into expression hosts.
Using Existing Hybridomas (for Recombinant mAbs)
Existing monoclonal antibodies that are hybridoma-based may be reproduced as recombinant monoclonal antibodies, resulting in specificity and consistency. This conversion process is accomplished by expressing the antibody-producing genes’ sequence obtained from the hybridoma in a mammalian cell line.
Using Phage Display (for Recombinant mAbs)
Through the use of in vitro phage display technology, it is possible to more quickly find recombinant monoclonal antibodies against targets, and also with no immunization of the animal. This process involves the production of an antibody library, the selection of antibodies that bind to the antigen/target of concern, and affinity maturation.