DNA Immunization Antibody Production

Creative Diagnostics offers proprietary genetic immunization based polyclonal and monoclonal antibody generation services. This unique antibody development approach involves direct immunization of host animals with plasmid DNA encoding the target protein of interest. The immunized hosts then produce the encoded protein and raise antibodies. Genetic immunization involves introducing the gene in the form of a cDNA directly into an animal which translates this cDNA into protein thus stimulating an immune response against the foreign protein. Protein purification is not necessary for this genetic immunization approach, which can save several months in time over recombinant protein generation followed by antibody production.

In order for genetic immunization to be successful, the cDNA-encoded protein must be secreted by the transfected cells in immunized animals or expressed on the surface of the transfected cells in order to get stimulation of an antibody response.

The foremost advantage of this antibody production approach is its high success rate in generation of high-affinity antibodies recognizing difficult-to-express proteins in their native confirmation, such as GPCRs, ion channels and other multiple membrane spanning proteins. For these proteins, recombinant protein fragments or peptides derived from their extracellular domains may raise antibodies workable in Western blotting but are extremely hard to produce high-affinity antibodies that can recognize their integral proteins in their native form. This point is important in raising antibodies for diagnostic use, in which recognition of the antigens in their native form can be required. For therapeutic antibodies, targeting the antigens in the native conformation with a high-affinity is of course required!

Of note, our technology allows guaranteed antibody development against 7-membrane-spanning GPCR proteins!

High affinity antibodies can result from genetic immunization because of low level of expressed proteins and constant presentation to the immune system; these tend to favor development of high affinity antibodies.

Usually polyclonal antibody development via genetic immunization is tried first since it is an economical way to see whether the protein-encoding plasmid/cDNA will raise the desired antibodies, e.g. antibodies recognizing an integral antigen in its native 3D conformation. If this is successful, monoclonal development is followed.