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Monoclonal Antibody Development

Monoclonal antibodies, following the development of this technology in 1975, have become an extraordinarily important resource for medical research, diagnosis, therapy, and basic science. They are antibodies which have a single, selected specificity and which, usually, are secreted continuously by “immortalized” hybridoma cells. A hybridoma is a biologically constructed hybrid of an antibody- producing lymphoid cell and a malignant (immortal) myeloma cell. The first step in the production of MAbs is to immunize a mouse with an antigen. When the mouse begins to produce antibodies to the antigen, its spleen is removed. Antibody-producing cells from the spleen are then fused with a myeloma cell line, one which is not antibody-producing and has been maintained in culture. The new fused cell line, which does produce antibodies, is grown briefly in culture and then re-injected into another mouse’s peritoneum. Finally, the ascites fluid which contains monoclonal antibodies is harvested from the mouse.

Millipore has developed and manufactured thousands of monoclonal antibodies. We provide custom hybridoma development, ascites production, cell culture production, cell cloning, hybridoma screening and clone banking. Our state of the art facilites are ISO13485,cGMP, and TGA certified. We offer small and large scale (multigram) production of monoclonal antibodies.

Custom Hybridoma Development Occurs in Phases as Follows

Millipore’s custom monoclonal antibody services include:
  • design concept development
  • antigen conjugation
  • cloning
  • screening
  • clone banking
Phase I: Immunization
Balb/c mice/rats or other rodent strain, are immunized with antigen provided by the client or developed by Millipore. Between five and ten animals will be immunized to optimize the probability of success.
Phase II: Cell Fusion
Cell fusion utilizes the spleenocytes from the hyperimmunized animals and an appropriate myeloma cell line (SP2/O, NS-1, or equivalent). Fusion is performed using PEG, and viable hybridoma colonies are grown out in 96 well plates.
Phase III: Screening & Selection of Clones
Cells are screened and selected by ELISA testing and/or Western blot. Up to five clones are selected. If additional clones exist, they will be made available for purchase.
Phase IV: Subcloning
Clones that are approved by the client are subcloned. Additional subclones will be made available for purchase.
Phase V: Colony Storage
Millipore will store the positive colonies that were not subcloned for a period of six months. Millipore can provide the client with additional colonies if desired. The remaining clones will be destroyed within 6 months or stored and banked per the client’s request.
Phase VI: Supernatant and Ascites Production
25 mLs of ascites will be provided for each clone. Hybridoma supernatant can be manufactured at an additional cost per clone, 500 mLs of supernatant is the standard volume provided.

The following quality control measures are employed to create the highest quality monclonal antibodies commercially available:
Standard Purity Checks Include
ELISA against the antigen
SDS/PAGE
IEF
HPLC
Western blot
Additional Functional Checks can be Performed per Client's Request Using
ELISA against the antigen
SDS/PAGE
IEF
HPLC
Western blot
For questions or comments please contact our specialized Custom Service Team at Millipore Corporation. Request a quote for Custom Services or Bulk Services

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