Phospho Glycan Lectin Array
Phosphorylation array is used for quick detection of a large number of site-specific phosphorylation of many kinases. Protein phosphorylation is a crucial cellular regulatory mechanism as numerous enzymes and receptors are activated or deactivated by phosphorylation and dephosphorylation events, by means of kinases and phosphatases. Capture antibodies are spotted on a nitrocellulose membrane and the levels of phosphorylated protein are assessed using biotinylated phospho-specific detection antibodies and visualized using the chemiluminescent detection reagent (Figure 1). Determining the phosphorylation of multiple kinases in a single sample can be expensive and time consuming, particularly if performing multiple immunoprecipitations and Western blots. The use of a multiplex antibody array for detecting multiple phosphorylation in a single sample can be cost-effective and also can save significant time and sample.
Figure 1. An illustration of RTK Phosphorylation Antibody Array assay.
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- Cui, Y., et al. Identification of the receptor tyrosine kinases (RTKs)-oriented functional targets of miR-206 by an antibody-based protein array. FEBS Lett. 2015, 589:2131-5. doi: 10.1016/j.febslet.2015.06.026.
Glycan array is used for profiling the interactions of glycan and glycan-binding proteins or partners in diverse biological samples in a high-throughput manner (Figure 2). Glycans are major components of the outermost surface of all animal and plant cells. In medical field, glycan profiling has aided research for identification of viral and bacterial infection, the inflammatory immune response, and progression of tumor cell metastasis, vaccine production, and medical diagnostics. APAF offers detection of binding partners for up to 300 different glycans in purified proteins, serum, plasma, cell culture supernatants, cell/tissue lysates or other body fluids.
Figure 2. An illustration of sandwich-based Glycan Array assay.
- Dan, X., et al. Development and Applications of Lectins as Biological Tools in Biomedical Research. Med. Res. Rev. 2016, 36:221-47. doi: 10.1002/med.21363.
- Krumm, S.A., and Doores, K.J. Targeting Glycans on Human Pathogens for Vaccine Design. Curr. Top. Microbiol. Immunol. 2018, 26. doi: 10.1007/82_2018_103.
- Trabbic, K,R., et al. Production of a mouse monoclonal IgM antibody that targets the carbohydrate Thomsen-nouveau cancer antigen resulting in in vivo and in vitro tumor killing. Cancer Immunol Immunother. 2018, 20. doi: 10.1007/s00262-018-2206-0.
Lectin array is a high-throughput glycan profiling technology where a panel of lectins are immobilized on a chip for assays (Figure 3). Lectins are glycan-binding proteins, purified from plants, which are highly specific for a given glycan based on their sequence and the different sugar unit in the glycan. Lectin array is used for studying glycosylation profiles of proteins, antibodies, cells, and cell lysates, and are applied in drug discovery and biomarker discovery studies. APAF offers detection of glycan profiles using up to 95 lectins; suitable for all liquid sample types.
Figure 3. An illustration of sandwich-based Lectin Array assay.
- Zhao, R., et al. Lectin array and glycogene expression analyses of ovarian cancer cell line A2780 and its cisplatin-resistant derivate cell line A2780-cp. Clin. Proteom. 2017, 14:20. doi: 10.1186/s12014-017-9155-z.
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