AI Article Synopsis
- The study aimed to measure the number of SV40 large tumor antigen (large T) molecules on the surface of SV40-transformed cell lines using cytofluorimetric analysis.
- Five different SV40-transformed cell lines were stained with a specific monoclonal antibody, and fluorescence signals were measured to determine the quantity of large T molecules per cell.
- Results showed varying amounts of large T molecules per cell across the different cell lines, highlighting the effectiveness of this technique for quantifying viral antigen expression on cell membranes.
Article Abstract
The aim of this study was to quantitate the number of cell membrane-located SV40 large tumor antigen (large T) molecules of SV40-transformed cell lines by cytofluorimetric analysis. Five different SV40-transformed cell lines were labelled by either a biotin- or a fluorescein-conjugated monoclonal antibody, PAb1605, which is specific for the large T carboxyterminus. The conjugated-antibody fluorescence signals of the stained large T molecules of transformed cells were measured via cytofluorimetry. Comparison of the fluorescence signals of calibrated beads bearing a known number of fluorescein molecules to the signals of conjugated PAb1605 antibodies bound on microbeads to a defined number of IgG binding sites made it possible to determine the number of antibody-accessible large T molecules per SV40-transformed cell. The numbers (x10(-4)) found per cell were 1.0 (ELONA, hamster), 3.0 (VLM, mouse), 3.5 (mKSA, mouse), 11 (C57SV, mouse), and 5.5 (SV80, human), respectively. Thus, the technique described allows a precise quantitation of surface-exposed, antibody-accessible viral antigen expression.
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| http://dx.doi.org/10.1002/cyto.990200112 | DOI Listing |
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