The glycan antigens of pathogens themselves represent a broad range of structures that incorporate diverse monosaccharides not typically found in the mammalian glycome. As a result, only limited information can be obtained following interrogation of host GBPs that recognize potential pathogens using conventional glycan microarray methods. In order to provide a more representative repertoire of the complex glycans in various pathogens, recent microarrays utilize glycans harvested directly from known microbes or synthetic microbial glycans in an array format, which generates a microbial glycan microarray (MGM). It composes of distinct and defined microbial glycans to interrogate the binding preferences and specificity of host immune factors with carbohydrate binding activity.
The general strategy of microbial glycan microarray analysis is similar to that of other types of microarray. The microbial glycans are either synthesized or isolated and purified from a broad range of microbes, and then undergo derivatization and printing in a microarray format. They are deposited on the NHS-activated glass slides over a wide range of concentrations to facilitate optimal detection of host-pathogen interactions. GBPs with putative pathogen recognition activity or sera are incubated with the microarray to examine potential interactions with microbial glycans. The detection is usually the fluorescence with a microarray scanner.
Figure 1. Schematic of microbial glycan microarray
MGM provides a unique platform of structurally diverse microbial glycans to interrogate the specificity of host factors directly involved in host pathogen interactions. It can accurately predict host-microbial interactions and can be interrogated rather easily to assess binding specificities of carbohydrate binding proteins, antibodies and even whole cells. MGM is sufficiently sensitive to pick up these changes of the adaptive immune systems in the presence of specific microbes. It is also able to uncover an entirely new innate immune mechanism for fighting off diverse bacteria that try to mimic mammals’ own cells.
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