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Hybridoma
What is a hybridoma?
A hybridoma is a cell arising from chemical fusion of two parental cell lines. One parent is an antibody secreting cell (usually from the spleen) isolated from an immunized animal. The other parent is a myeloma cell (a type of B cell tumor). Hybridomas are immortal somatic cell hybrids that secrete antibodies.
What are monoclonal antibodies?
Monoclonal antibodies are produced (secreted) by cloned populations of hybridoma cells.
What are polyclonal antibodies?
Polyclonal antibodies are isolated from the serum of an immunized animal. There are typically thousands of different antibodies reacting with numerous epitopes and affinities present in polyclonal antibody preparations.
Of what use are hybridomas and monoclonal antibodies?
Monoclonal antibodies produced by hybridomas are used in many areas of basic scientific research, industry, human and animal medicine and agriculture. Hybridomas can be generated that secrete antibodies to almost any substance (toxins, drugs, blood proteins, cancer cells, viruses, hormones, environmental pollutants, food products, metals, plant materials). The list of substances to which monoclonal antibodies have been made is almost endless. Monoclonal antibodies are routinely used to create sensitive tests for detecting or quantitating the presence or amount of various substances. Scores of monoclonal antibody based tests are used in human and animal clinical labs. Monoclonal antibodies can be used to isolate and purify specific compounds from complex mixtures (immunoaffinity chromatography). Monoclonal antibodies are also used therapeutically to treat diseases such as cancer.
Do I need monoclonal or polyclonal antibodies for my application?
For many applications, such as electron microscopy, polyclonal antibodies may offer some important advantages over monoclonal antibodies. Choosing whether to use a polyclonal or monoclonal antibody for a certain technique requires careful consideration. Often a pool of well defined monoclonal antibodies can accomplish many of the same things as a polyclonal reagent.
Does the Hybridoma Core Lab make polyclonal antibodies?
No. We can provide contact information for a number of commercial vendors who provide this service.
How much antigen do I need to generate a monoclonal antibody?
The amount of antigen needed to generate a monoclonal antibody varies greatly from one antigen to another. Projects involving antigens that are extremely immunogenic (such as insect lectins) may only require 0.3-0.6 mgs. Projects where the antigen is weakly immunogenic (such as short peptides) may require 1.0 - 5.0 mgs. General considerations that come into play when estimating amount of antigen needed include: number of mice to be immunized, dose required to generate an immune response, amount of antigen needed for performing assays on mouse serum and hybridoma supernatants.
How can I find out if a certain antibody is commercially available?
Follow the How to find an antibody link on the Antibody Resource page on the internet: http://antibodyresource.com/. The Linscott's Directory of Immunologogical and Biological Reagents, which is a compilation of many commercially available reagents and custom services is available for free online searching: http://www.linscottsdirectory.com/
How long does it take to generate a monoclonal antibody?
Development of a monoclonal antibody usually takes from 4-6 months. The time frame will vary depending on how many immunizations are required to generate an appropriate immune response in the mice.
Do I need to obtain my own Animal Use Approval number?
Yes, the University of Florida Institutional Animal Care and Use Committee requires that each investigator that contracts for work involving animals fill out an Animal Use Approval Form. If your only requirement for animal use is for monoclonal antibody production by the Hybridoma Core Lab, there is an abbreviated form you can download from: http://iacuc.ufl.edu/Forms/hybridomaform.doc. This site has instructions on how to submit the completed form for approval.
How much does it cost to have a monoclonal antibody made?
For the Hybridoma Core Lab to do a complete project starting with immunizing a group of mice with an antigen, screening test bleeds for antibody response, performing one cell fusion, assaying the fusion by ELISA, providing supernatants to the investigator lab for secondary screening, cloning one or two cell lines and producing a monoclonal antibody from the best clone, the cost estimate is $2,500.00 - $3,000.00 for University of Florida investigators. Our laboratory is subsidized by the University of Florida which pays for staff salaries, instrumentation and maintenance. Our fees (UF rate) recover the cost of supplies and reagents used for each job.
What things should I consider before beginning a monoclonal antibody project?
There are several things to carefully think about before beginning a monoclonal project, such as: Does a monoclonal or polyclonal antibody already exist (from commercial sources or from a colleague) that may work for my application? Will a monoclonal or polyclonal antibody work best for my needs? Do I have enough antigen prepared to complete the project? Can I set up a relevant assay to lead to monoclonal antibodies that will suit my needs?
How do I start a monoclonal antibody project with the Hybridoma Core Lab?
Contact Linda Green by email or call to arrange a brief face to face or phone conference to discuss your needs and the project details.

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Electron Microscopy & Bioimaging
How do I sign up to use an EMBL instrument?
To sign up to use an EMBL instrument, simple click on the link below that corresponds to the equipment you wish to use.

 
 

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