PEGS Europe is the largest European event covering all aspects of protein and antibody engineering. Over the last three years, participation at PEGS Europe has grown by more than 60%, and this year will be the largest ever. Takis has participated at the last three editions creating a successful network.
This year, Dr Giuseppe Roscilli will present a paper entitled "IMMUNE LIBRARY AND ANTIBODY GENERATION TO DIFFICULT TARGETS BY ELECTRO-GENE-TRANSFER".
It is well known that classical antibody generation is limited by the availability of properly-folded and purified antigen and Takis genetic immunization technology represent powerful tool to induce immune response against soluble or membrane expressed antigens. We routinely utilized DNA electro-gene-transfer (DNA-EGT) to generate monoclonal antibodies to different antigens without the need to purify the protein. This technology is able to induce strong humoral response in mice when selected peptides or proteins are completely ineffective. Above all, DNA-EGT is able to break tolerance against self antigens. Moreover engineering, such as codon-optimization and fusion with immunoenhancing moieties and combinations with immunomodulators have a significant impact on antibody titer and relative isotype.
We have introduced this technique in a typical workflow to generate a set of novel hybridomas but skipping the classical somatic mutation. From immunized animal B cells an immune library is obtained by PCR amplifing VH and VL and cloning them in an eukaryotic expression vector expressing constant regions of the human H and L IgG genes. Antibiotic resistant gene expressed by the vector will allow the selection of transfected cells and expression of the desired antibody can be assessed by classical ELISA or a functional assay. The clone expressing the antibody with the desired characteristics can be sequenced, subjected to affinity maturation or directly subcloned and expanded to produce the antibody of interest.
This workflow represents a faster and more efficient approach than conventional somatic fusion for the identification of a novel antibody in a chimeric format expressed in a recombinant clonal cell line.