Hybridoma technology steps for mAbs production

Our hybridoma generation service is based on a stepwise approach.

We generate antibody producing clones from a large panel of antigens.
We send reports to our customers at each phase of the monoclonal antibody production process.

We remain available all along the program to discuss potential adaptations with our customers.
If no positive hybridoma is obtained during phase 3, only phase 1 will be charged.

 

 

 

Set-up

  1. Project discussion

    Selection of the best strategy

  2. Antigen design and production

    Various antigens can be used:

    • peptides
    • proteins
    • bacteria
    • inactivated viruses
    • transfected cells
    • small molecules

    Others – please discuss your antigen needs with our specialists

Hybridoma Development Phases

  1. Murine Immunization

    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.

  2. Fusion

    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

  3. 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.

  4. 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-lasting experience in monoclonal antibody production from hybridoma cultured in vitro.

We can produce up to 100g scale mAbs per batch.

Continue to mAb production

Option for your monoclonal program

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.