Monoclonal Antibodies South Australia (MAbSA) Technologies

Facility Managing Scientist:
Dr. Alfio Comis
Telephone: +61 8 8261 1033
Facsimile: +61 8 8261 2280
Mobile: +61 0435 658 435
alfio.comis@imvs.sa.gov.au

IMVS
Veterinary Services Division

101 Blacks Road
Gilles Plains, SA, 5086
Australia

Monoclonal Antibodies SA (MAbSA) is a state-of-the-art facility for the development of hybridoma cell lines and the purification of monoclonal antibodies. MAbSA is the only facility of its kind in South Australia. Known for its cost-effective and high quality service, MAbSA has clients throughout Australia and overseas.

MAbSA offers a wide variety of tissue culture services that are designed to produce an antibody product that is the perfect reagent for your applications. This includes antigenic peptide design, hybridoma production, antibody harvesting, affinity purification, conjugation, fragmentation and immunoassay development. We have made antibodies to large and small peptides, proteins, organic molecules, and bacterial as well as viral antigens.

All queries are promptly handled in a professional manner. We understand and appreciate that every project is unique and our hybridoma development programs are always tailored to the needs of the researcher. Therefore, we analyse and discuss all the important features of the antigens and the antibody characteristics required by each project. During the course of the project, we will keep the client up to date with regular email or telephone progress reports. The client is free to perform any part of the project and engage MAbSA to do the rest. For example, the client may choose to do the immunizations and provide MAbSA with a mouse ready to be used for the fusion. Or the client may choose to clone its own cell line after MAbSA has identified all the positive populations after fusion. We are as flexible as the client wants us to be.

Organization of each project:

Project are tailored to the need of the client and his/her project.
Typical completion time is 5-6 months.
Our programs are made of separate components and the client can choose any of the modules to be carried out by MAbSA and the rest to be left for the client's own laboratory to complete.
All positive supernatant samples are made available to the client for testing in the most appropriate system before selecting the best cell line for subcloning.
Affinity-purified antibody can be produced in any quantity from each clone selected by the client.
All cell lines and antibodies are the property of the client and are kept at two separate locations until directed otherwise by the client.
Each project is made up by separate components for billing purpose. Each phase is billed at completion and before the next phase is started, so that only successful phases are charged.

Synthetic peptides can be cross-linked to carrier proteins for immunization and/or for screening purpose.
Peptides can be cross-linked through amino, carboxyl, or thiol groups depending on the peptide.
Other chemistries are also available on request.
Other haptens can also be cross-linked to carrier proteins depending on the chemical structure.

Immunization of 3 Balb/c mice. More animals can be used if required.
Immunogen is injected at three week intervals to increase the chance of producing higher affinity antibody responses.
Test bleeds are evaluated by ELISA and sent to the client for testing if so required.
The animal with the highest titre is initially selected for fusion.
If required, each animal will be used for fusion until a good positive antibody suited for the job is found.

Spleen cells from selected animals are fused with one of two available myeloma cell lines (NS/1 or SP2/0).
All clones originated are screened by ELISA and all positive clones are frozen down for future use, if required.
Supernatant from positive clones are sent to client for assessment in the most appropriate system.
The client chooses any cell line required for cloning.

Each hybridoma chosen by the client is subcloned to obtain a stable monoclonal-antibody secreting cell line.
Liquid supernatant samples are available to the clients at each phase of the subcloning process if required for evaluation in their systems.

The selected hybridoma is weaned to grow in the best of a number of serum-free media available to MAbSA.
Cells are then grown in large quantities and harvests are collected every week. Each harvest of about 15 ml will yield on average 10 mg of antibody after purification.
The harvests are purified through a protein G affinity column and the product extensively dialysed and freeze-dried.
The amount of antibody produced can be increased by putting on line more reactors.

Estimated time for producing an average of 30mg of affinity-purified antibody is 4-6 weeks for each reactor.

MAbSA can subclone hybridomas with diminished levels of antibody production to establish new monoclonal cell lines selected for maximum antibody productivity.
We will also subclone hybridomas when monoclonality has been shown to be compromised.
All new subclones will be tested for antibody production, or samples can be sent to the client for testing.

MAbSA can perform a new fusion of any hybridoma that no longer grows efficiently or any hybridoma with diminished level of antibody production to a different myeloma cell line to establish new monoclonal cell lines. All new subclones will be tested for antibody production, or samples can be sent to the client for testing.
Each hybridoma chosen is subcloned to obtain a stable monoclonal-antibody secreting cell line.
Supernatant fluid samples are available to the client at each phase of the subcloning process if required for evaluation in their systems.
Stable cell line is delivered to the client either frozen or in culture.

MAbSA will prepare frozen stocks of cells by culturing sufficient cell numbers to be suspended in cell freezing medium and transferred to 8 cryogenic vials (at least 1 million cells per vial).
Vials of cells are taken through a careful freezing process and, finally, frozen in liquid nitrogen tanks.
MAbSA will ship the cells on dry ice or in culture.
Cell stocks will be kept at two geographically separate locations.
One vial of cells will be reconstituted on request to confirm cell viability.
Cell lines can also be submitted to MAbSA for storage and they must have been tested for Mycoplasma and be documented to be negative.