SERVICES

Any researcher requesting information regarding the services available from the Antibody Core Facility is encouraged to contact:

507-284-4701

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Project Consultation:

Prior to initiation of a project, a mutually acceptable research strategy is outlined. Antigen preparation, immunization protocols, supernate screening strategies, characteristics of desired monoclonal antibodies and individual responsibilites are established before work is undertaken.

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Mice:

Mayo Foundation’s Institutional Animal Care and Use Committee (IACUC) will review prospective monoclonal antibody projects. Request for animal authorization is submitted in writing to IACUC via an IACUC addendum form. The IACUC addendum submission forms can be obtained by contacting the Mayo Antibody Core Facility.

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Immunizations and Antisera Collection:

Generally, five mice are immunized, boosted and individual bleedings are collected to identify animals producing antibody of interest. Cell hybridizations are performed using one donor spleen/fusion.

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Fusions:

Cell hybridizations are performed using the FO cell line. FO is a HAT sensitive, immunoglobulin nonsecreting, mouse myeloma. Ten 96 microtiter plates are seeded per fusion. Fusion efficiencies vary and an investigator can anticipate screening anywhere from 50 to 960 primary supernates.

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Screening:

Mayo Antibody Core Facility will perform an initial screening utilizing only ELISA assays. Other screening assays (e.g. Western blotting, immunoprecipitation, immunohistochemistry) are the responsibility of the client.

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Hybridoma Cloning and Subcloning:

Hybridomas of interest are subjected to two rounds of cloning in order to insure monoclonality and stability. The Mayo Antibody Core Facility can perform ELISA assays on clone and subclone supernates. Other assays remain the responsibility of the client.

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Isotyping:

Isotyping of isolated monoclonal antibodies can be performed by the Mayo Antibody Core Facility.

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Cryopreservation:

Six vials of each monoclonal antibody clone are cryopreserved in liquid nitrogen. A fee will be assessed for long term storage of vials.

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Scale-up of Antibody Production:

The Antibody Core Facility does not produce ascites fluid as a means of generating larger amounts of monoclonal antibodies. Larger amounts of desired monoclonal antibodies are produced in roller bottles. Yields of monoclonal antibodies in roller bottles are clone dependent and vary from 5-50 mg/roller bottle.

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Antibody Purification:

Monoclonal antibodies are isolated from culture supernates by affinity chromatography using Protein G columns.

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Polyclonal Antisera :

Under some circumstances larger amounts of polyclonal sera can be produced contingent upon IACUC approval.