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National Repository of Grey Literature 2 records found  Search took 0.01 seconds. 
Preparation of monoclonal antibody against the major structural protein, VP1, of BK virus and construction of plasmids for protein interaction analysis by BiFC method.
 Kozmanová, Alexandra ; Forstová, Jitka (advisor) ; Němečková, Šárka (referee)
The aim of this study was to create a monoclonal antibody against VP1 protein BK virus for study of individual steps of its replication cycle. BK virus is human polyomavirus which causes serious disease - polyomavirus associated nephropathy in immunocompromised patients after renal transplantation or hemorrhagic cystitis after bone-marrow transplantation. Virions of BK virus have capsids with icosahedral symmetry consisting of 360 molecules of the major capsid protein VP1 and two minor structure proteins VP2 and VP3, which are not exposed on the capsid surface. VP1 is highly immunogenic what gives us possibility to produce antibodies capable of BK virus monitoring by capsid protein detection. Empty BK virus particles produced in insect cells from a recombinant baculovirus were used as antigen to immunize mice for preparing hybridomas. Cloning of hybridomas created by fusion of myeloma cells with B-lymphocytes from immunized mouse gave us series of primary hybridoma clones. One of them, 5/G10/A5/B2/8, was cloned to homogenity and monoclonal antibody against VP1 BK virus was purified from hybridomas medium by affinity chromatography. The analysis of antibody properties revealed that it recognizes a conformation epitope and can be used for VP1 protein and virion detection in cells by indirect...
Detailed record
Preparation of monoclonal antibody against the major structural protein, VP1, of BK virus and construction of plasmids for protein interaction analysis by BiFC method.
 Kozmanová, Alexandra ; Forstová, Jitka (advisor) ; Němečková, Šárka (referee)
The aim of this study was to create a monoclonal antibody against VP1 protein BK virus for study of individual steps of its replication cycle. BK virus is human polyomavirus which causes serious disease - polyomavirus associated nephropathy in immunocompromised patients after renal transplantation or hemorrhagic cystitis after bone-marrow transplantation. Virions of BK virus have capsids with icosahedral symmetry consisting of 360 molecules of the major capsid protein VP1 and two minor structure proteins VP2 and VP3, which are not exposed on the capsid surface. VP1 is highly immunogenic what gives us possibility to produce antibodies capable of BK virus monitoring by capsid protein detection. Empty BK virus particles produced in insect cells from a recombinant baculovirus were used as antigen to immunize mice for preparing hybridomas. Cloning of hybridomas created by fusion of myeloma cells with B-lymphocytes from immunized mouse gave us series of primary hybridoma clones. One of them, 5/G10/A5/B2/8, was cloned to homogenity and monoclonal antibody against VP1 BK virus was purified from hybridomas medium by affinity chromatography. The analysis of antibody properties revealed that it recognizes a conformation epitope and can be used for VP1 protein and virion detection in cells by indirect...
Detailed record

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