We generate murine hybridomas
from a large panel of antigens using a stepwise approach
A report is sent after each phase and may be discussed with the customer for potential adaptation.
If no positive hybridoma is obtained during phase 3, only phase I is charged.
Selection of the best strategy
Antigen design and production
Various antigens can be used:
- inactivated viruses
- transfected cells
- small molecules
Others – please discuss your antigen needs with our specialists
The success highly depends on the antigen quality. By default, four mice are immunized to allow selecting of the best responder. Immune responses are evaluated by ELISA.
Splenocytes from one mouse whose immune response was found positive in ELISA are fused with a murine myeloma cell line.
Using the GentleMACS and optimizing the myeloma bank management make this fusion step a very reproducible process
Hybridoma screening & selection1
Please note that the screening step is very critical as clones in culture are short-lived. Hence the screening strategy must be defined in advance and prompt reactivity will be mandatory to consider the customer’s input.
The project will be considered as successful if positive clones are obtained.
Cloning, isotyping and cell banking
In Phase 3, screening usually happens on hybridoma mixtures, part of which only being secreting the desired mAb.
As the clones selected are not yet monoclonals, they are further expanded and subcloned during Phase 4 to guaranty monoclonality. Isotypes are determined, and 5 ampules per clone are stored in liquid nitrogen.
Want to produce mAb
from your hybridoma?
We have a long expertise in producing small up to very large scale mAbs exclusively in vitro.
1 The screening step is by default performed in ELISA, and clones will be considered positive if they generate a signal against the immunogen. We guaranty that the mAbs selected from this screening step will recognize the antigen in ELISA; but we cannot guaranty that they will recognize and bind to any other form of the antigen not used during the immunization or screening. The project will be considered as successful if positive clones are obtained.
The screening step is very critical as clones in culture are short-lived. Hence the time to screen is very limited, and the screening strategy must be defined in advance. Clone supernatants can be shipped to the customer for on-site analyses, but prompt reactivity will be mandatory to consider the customer’s input for the clones’ selection. Depending on the final application, the screening phase can be performed using several technologies besides ELISA, including FACS, WB and Dot Blot.