title: New sensitive detection method for lectin hemagglutination using microscopy.
abstract: The blood group system AB0 is determined by the composition of terminal oligosaccharides on red blood cells. Thanks to this structural feature, these groups can be recognized by saccharide-recognizing compounds. Lectins are proteins that are able to reversibly bind saccharide structures. They generally occur as multimers and are known as hemagglutination agents. Hemagglutination is a process in which blood cells are cross-linked via multivalent molecules. Apart from lectins, hemagglutination can also be caused by antibodies or viruses. A hemagglutination assay is commonly used for the detection of multivalent molecules that recognize blood cells, in order to search for their sugar specificity. It is traditionally performed on a microtiter plate, where the lectin solution is serially diluted and the lowest concentration of lectin causing agglutination is detected. This experimental set-up is utilized further for testing lectin specificity via a hemagglutination inhibition assay. We have developed a new way of detecting hemagglutination using microscopy, which was tested on purified lectins as well as cell lysates. Hemagglutination was performed on a microscope slide directly and detected using a microscope. Comparison with the standard hemagglutination assay using microtiter plates revealed that microscopic approach is faster and more robust and allows fast determination of lectin activities immediately in bacterial cytosols.
authors: Central European Institute of Technology, Masaryk University, Kamenice 5, 625 00, Brno, Czech Republic; National Centre for Biomolecular Research, Faculty of Science, Masaryk University, Kotlářská 2, 611 37, Brno, Czech Republic.
authors: Lenka
authors: L
authors: Adamová
authors: Lenka
authors: L
authors: Malinovská
authors: Michaela
authors: M
authors: Wimmerová
mesh_terms: D006384:Hemagglutination; D006385:Hemagglutination Inhibition Tests / Q000379:methods; D006386:Hemagglutination Tests / Q000379:methods; D006461:Hemolysis; D006801:Humans; D037102:Lectins / Q000378:metabolism*; D008853:Microscopy / Q000379:methods
chemical_list: D037102:Lectins
keywords: blood group antigens; hemagglutination; lectin; protein-carbohydrate interactions