New sensitive detection method for lectin hemagglutination using microscopy.

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Authors

ADAMOVÁ Lenka MALINOVSKÁ Lenka WIMMEROVÁ Michaela

Year of publication 2014
Type Article in Periodical
Magazine / Source Microscopy research and technique
MU Faculty or unit

Central European Institute of Technology

Citation
Web http://dx.doi.org/10.1002/jemt.22407
Doi http://dx.doi.org/10.1002/jemt.22407
Field Biochemistry
Keywords hemagglutination; protein-carbohydrate interactions; blood group antigens; lectin
Description 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. Hemagglutination is a process in which blood cells are cross-linked via multivalent molecules, lectins e.g. 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. 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.
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