Reusable glycan microarrays using a microwave assisted wet-erase (MAWE) process

Author:

Mehta Akul Y12,Tilton Catherine A12,Muerner Lukas1234,von Gunten Stephan5,Heimburg-Molinaro Jamie12,Cummings Richard D12

Affiliation:

1. Department of Surgery , Beth Israel Deaconess Medical Center, National Center for Functional Glycomics, , 3 Blackfan Circle, Center for Life Sciences, Boston, MA 02115 , United States

2. Harvard Medical School , Beth Israel Deaconess Medical Center, National Center for Functional Glycomics, , 3 Blackfan Circle, Center for Life Sciences, Boston, MA 02115 , United States

3. Institute of Pharmacology, University of Bern , , Switzerland

4. Inselspital, INO-F, Bern 3010 , , Switzerland

5. Institute of Pharmacology, University of Bern , Inselspital, INO-F, Bern 3010 , Switzerland

Abstract

Abstract Modern studies on binding of proteins to glycans commonly involve the use of synthetic glycans and their derivatives in which a small amount of the material is covalently printed onto a functionalized slide in a glycan microarray format. While incredibly useful to explore binding interactions with many types of samples, the common techniques involve drying the slides, which leads to irreversible association of the protein to the spots on slides to which they bound, thus limiting a microarray to a single use. We have developed a new technique which we term Microwave Assisted Wet-Erase (MAWE) glycan microarrays. In this approach we image the slides under wet conditions to acquire the data, after which the slides are cleaned of binding proteins by treatment with a denaturing SDS solution along with microwave treatment. Slides cleaned in this way can be reused multiple times, and an example here shows the reuse of a single array 15 times. We also demonstrate that this method can be used for a single-array per slide or multi-array per slide platforms. Importantly, the results obtained using this technique for a variety of lectins sequentially applied to a single array, are concordant to those obtained via the classical dry approaches on multiple slides. We also demonstrate that MAWE can be used for different types of samples, such as serum for antibody binding, and whole cells, such as yeast. This technique will greatly conserve precious glycans and prolong the use of existing and new glycan microarrays.

Funder

National Institutes of Health

Massachusetts Life Sciences Center

Swiss National Science Foundation

University of Bern

Publisher

Oxford University Press (OUP)

Subject

Biochemistry

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