- The Monoclonal Antibody Production process requires several months.
The General Procedure Includes the Following Monoclonal Antibody Development Steps:
- Immunization of mice with the antigen of interest.
- Screening of mice antibody titer using ELISA. This step can tell if animals' immune systems responded to the antigens injected. It also indicates which animal has the highest response.
- Thawing and preparation of myeloma cells for monoclonal antibody production.
- Preparation of single cell suspension from mice spleens.
- Fusion of antibody-producing B cells from spleens and myeloma cells.
- Selective growth of the hybridoma in HAT.
- Screening and sorting the hybridomas for producing the desired monoclonal antibody using FACS.
- Cloning of selected hybridomas.
- Screening of hybridomas for antibody production using ELISA.
- Freezing of the hybridoma.
The Monoclonal Antibody Manufacturing Process:
- The first step is Immunization, where a mouse is injected with the antigen, to which an antibody is to be generated.
- The rat’s immune system responds to the antigen by producing B cells. Each B cell produces a single type of antibody.
- B cells are isolated from the mouse’s spleen and fused with immortal myeloma cells producing ‘hybridoma cells’ through hybridoma technology and monoclonal antibodies production.
- This fusion is done with the help of polyethylene glycol (PEG).
- Hybridomas should be able to produce the desired antibody through monoclonal antibodies hybridoma technology. They are sorted using flow cytometry (FACS).
Thus; hybridomas can be cloned into a 96 well plate in a 1cell/well ratio. When colonies become confluent, they can be screened for antibody production using ELISA in the monoclonal antibody production workflow.
