Production, characterization and application of new monoclonal antibodies against viral antigens

Indrė Kučinskaitė-Kodzė

Doctoral dissertation

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Technological sciences, chemical engineering (05T), Biotechnology (T490).

The dissertation describes development and characterization of monoclonal antibodies against recombinant yeast-expressed antigens: nucleocapsid (N) proteins of human parainfluenza virus type 3, Menangle virus, hantavirus and rabies virus. The newly developed antibodies were investigated by different immunochemical assays for their specificity, affinity and ability to recognize native viruses in infected cells. It was determined that the antibodies raised against recombinant yeast-expressed viral proteins are suitable to identify virusinfected cells. These data confirmed that recombinant yeast-expressed viral N proteins possess antigenic properties similar to that of native viral nucleocapsids. The monoclonal antibodies were also used to study the antigenic structure of viral N proteins and localize their immunodominant regions. The obtained results may have impact on the development of new immunodiagnostic test systems for the detection of viral infections.

The dissertation consists of the introduction, three sections, references, and the list of author’s publications.

In the Introductory Chapter, the research topic, the actuality, the aim and tasks, scientific novelty and practical value of the dissertation are discussed. Author’s publications and conference reports are also presented. The first Chapter of the dissertation provides literature overview on the genome organization, structural proteins, pathogenesis and epidemiology of parainfluenza viruses, Menangle virus, hantaviruses and rabies viruses. The properties of viral N proteins are described in more detail. The literature overview also demonstrates the benefits of the expression of recombinant viral proteins in yeast, describes commonly used diagnostic assays for viral infections and application of monoclonal antibodies for the immunodiagnostics of viral infections. In the second Chapter, materials and methods are presented: hybridoma technology, enzyme-linked immunosorbent assay (ELISA), Western blot, immunofluorescence microscopy, molecular biology techniques. The third Chapter provides experimental data on the development of new monoclonal antibodies, testing of their specificity and reactivity with different viral N proteins, infected cells and tissues, the use of monoclonal antibodies for studying the antigenic structure of viral N proteins.

Four articles have been published on the subject of the dissertation. All publications have been included into ISI Web of Science list. The results of the dissertation have been presented in 6 scientific conferences in Lithuania and abroad.

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Imprint No:
160×230 mm
146 p.
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