"DNA To Antibody": Genetic Immunization
QED Bioscience is proud to be the first U.S. antibody company to offer custom monoclonal and polyclonal antibody development through genetic immunization. This powerful approach to antibody development involves the direct immunization of the host animal with plasmid DNA encoding the client's protein of interest. The immunized host then produces the encoded protein and raises antibodies. Genetic immunization is perfectly suited to producing antibodies when a protein is difficult to express, purify or when a gene has been obtained, but the protein itself is unknown.
Features and Benefits:
- No additional charge when used with either monoclonal or polyclonal antibody development programs for Genetic Immunization.
- Save Time: Large-scale synthesis and purification of proteins are no longer necessary.
- Control Your Target: Antibodies are directed only against the encoded protein. Unwanted antibodies to contaminants found in purified protein preparations are eliminated.
- Produce High Affinity Antibodies: Much lower amounts of protein generated by genetic immunization, as well as slow, consistent presentation to the immune system, favor the production of high affinity antibodies. Plasmid DNA also acts as an adjuvant to stimulate immune responses.
- Customize Antibodies To Your Applications: These antibodies can be used in many applications, including, but not limited to, purifying proteins from cells or tissues, identifying proteins in prognostic and diagnostic assays, etc.
The following guidelines are provided to assist you in preparing plasmid DNA for QED's genetic immunization programs:
- Choice Of Expression Vector
The plasmids pcDNA3.1 containing the CMV promoter and pRc/RSV from Invitrogen have been used successfully. In unusual cases where over-expression of a protein is cytotoxic, a weaker promoter (such as SV40) is a better choice.
- Confirmation Of Protein Production
It is recommended that you perform transient transfection of a mammalian cell line (such as COS, 3T3, or muscle cell lines) in order to confirm that your DNA-encoded protein can be produced in mammalian cells.
- Preparation Of DNA For Immunization
Plasmid DNA should be dissolved in phosphate-buffered saline at a concentration of approximately 1-2 mg/ml. A dose of 100 ug/injection/animal will be administered. We expect to administer 3-4 injections/animal, two weeks apart.
- Evaluation Of Immunized Animals
A small amount of your DNA-encoded protein (~1 mg) is required for testing antibody responses of immunized animals. QED will test serum antibody titers in ELISA unless you require a different assay format. Of course, QED will send serum samples from immunized animals to you if you prefer to do this testing in your laboratory.
- Suitability of the Antigen: Three criteria must be met in order for your antigen to be suitable for this procedure:
- The protein must be expressed by the plasmid.
- Once the protein is expressed in vivo, it must be made available to the immune system. Proteins that are exported from the cell or placed on the cell membrane are suitable; nuclear proteins, for example, are not.
- The protein, upon presentation to the immune system, must be antigenic. Be sure to check the sequence homology of your protein to that of the host.
Please let us know if you have any questions about these Guidelines. We look forward to working with you!
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