Dipodal Silanes Greatly Stabilize Glass Surface Functionalization for DNA Microarray Synthesis and High-Throughput Biological Assays
- Author(s)
- Arya Das, Santra Santhosh, Maya Giridhar, Jürgen Behr, Timm Michel, Erika Schaudy, Glenda Gisela Ibanez Redin, Jory Lietard, Mark M Somoza
- Abstract
Glass is by far the most common substrate for biomolecular arrays, including high-throughput sequencing flow cells and microarrays. The native glass hydroxyl surface is modified by using silane chemistry to provide appropriate functional groups and reactivities for either in situ synthesis or surface immobilization of biologically or chemically synthesized biomolecules. These arrays, typically of oligonucleotides or peptides, are then subjected to long incubation times in warm aqueous buffers prior to fluorescence readout. Under these conditions, the siloxy bonds to the glass are susceptible to hydrolysis, resulting in significant loss of biomolecules and concomitant loss of signal from the assay. Here, we demonstrate that functionalization of glass surfaces with dipodal silanes results in greatly improved stability compared to equivalent functionalization with standard monopodal silanes. Using photolithographic in situ synthesis of DNA, we show that dipodal silanes are compatible with phosphoramidite chemistry and that hybridization performed on the resulting arrays provides greatly improved signal and signal-to-noise ratios compared with surfaces functionalized with monopodal silanes.
- Organisation(s)
- Department of Inorganic Chemistry
- External organisation(s)
- Technische Universität München
- Journal
- Analytical Chemistry
- Volume
- 95
- Pages
- 15384-15393
- No. of pages
- 10
- ISSN
- 0003-2700
- DOI
- https://doi.org/10.1021/acs.analchem.3c03399
- Publication date
- 2023
- Peer reviewed
- Yes
- Austrian Fields of Science 2012
- 104004 Chemical biology, 104002 Analytical chemistry
- Keywords
- ASJC Scopus subject areas
- Analytical Chemistry
- Portal url
- https://ucrisportal.univie.ac.at/en/publications/0767720a-bcb3-4d7d-ab49-97fc2fa3dee4