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Establishment of high-throughput monoclonal antibody production and hybridoma bank

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New bank for monoclonal antibodies

An EU-funded project CAPRI has investigated what is needed to upgrade research on monoclonal antibodies and make the information available to all scientists in the field.


Research has produced many monoclonal antibodies (mAbs) against thousands of proteins that may be used for diagnostic tests to detect substances, cancer treatment for an immunological response against the target cancer cell and in autoimmune disease treatment to inhibit the inflammatory response. Hybrid cell lines can also be produced by fusing together a specific antibody-producing B cell with a cancer cell. The scope of therapeutic use for mAbs is therefore extremely wide and the number of different species for the treatment of viruses, cancer and immune system disorders as well as substance identification would seem almost infinite. In view of the demand for these potential molecular miracles, researchers with the EU-funded project CAPRI established a research and biotechnology unit specifically for production of mAbs together with a hybridoma and mAbs bank. The first goal achieved was to generate mAbs against proteins of all relevant human viruses starting with the entire proteome of the Variccela zoster virus (VZV), the cause of human chickenpox and shingles. The basic screening was performed using ELISA followed by Western blot (WB) on the protein produced in Escherichia coli (E. coli). Confocal microscopy and WB were used for cells infected with VZV. To prevent the production of mAbs that fail to react to their natural antigens from VZV-infected cells, the scientists used two different strategies. First, the fragmentation of the original open reading frames (ORFs) where there is no stop codon and therefore may be used to produce a protein. This avoids potential hydrophobic and transmembrane regions. Secondly, the team established a baculovirus expression system; the resulting Baculovirus lacking one of the non-essential genes replaced with a foreign gene. As well as the research initiatives, a hands-on approach was organised for all researchers interested in mAbs. A summer school on 'Innate Immunity to viral infection', meetings to discuss potential collaborations and individual tuition in mAbs techniques at the University of Rijeka have encouraged interaction between experts in the field. Online, a database with laboratory management software 'webLAB' provides detailed information about proteins, hybridomas and mAbs and is updated on a regular basis. Overall, the impressive results of the CAPRI initiative will contribute to the upgrading of resources dedicated to the application of genomics and biotechnology for health purposes.

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