Carbohydrate Microarrays by Microcontact Printing

Abstract

This Article describes the preparation of carbohydrate microarrays by the immobilization of carbohydrates via microcontact printing (microCP) on glass and silicon substrates. To this end, diene-modified carbohydrates (galactose, glucose, mannose, lactose, and maltose) were printed on maleimide-terminated self-assembled monolayers (SAMs). A Diels-Alder reaction occurred exclusively in the contact area between stamp and substrate and resulted in a carbohydrate pattern on the substrate. It was found that cyclopentadiene-functionalized carbohydrates could be printed within minutes at room temperature, whereas furan-functionalized carbohydrates required long printing times and high temperatures. By successive printing, microstructured arrays of up to three different carbohydrates could be produced. Immobilization and patterning of the carbohydrates on the surfaces was investigated with contact angle measurements, X-ray photoelectron spectroscopy (XPS), time-of-flight secondary ion mass spectrometry (TOF-SIMS), and fluorescence microscopy. Furthermore, the lectins concanavalin A (ConA) and peanut agglutinin (PNA) bind to the microarrays, and the printed carbohydrates retain their characteristic selectivity toward these proteins.

Molecular recognition of surface-immobilized carbohydrates by a synthetic lectin

“…Lectin carbohydrate interactions: As described in [34], in order to reduce non-specific protein adsorption, the arrays were incubated with a 3% bovine serum albumin (BSA) solution in PBS buffer (pH 7.5, 0.1% Tween 20) for 30 min and washed two times with PBS buffer prior to lectin screening. The surfaces of the carbohydrate arrays were covered by a solution of 1 mM FITC HisHis and 1 μg of labeled lectin (TRITC ConA or FITC PNA) in 100 μL of HEPES buffer (20 mM HEPES, pH 7.5, 0.15 M NaCl, 1.0 mM CaCl 2 ).…”

“…Our group has shown that also simple carbohydrate microarrays can be conveniently prepared by μCP if reactive glycosides are printed on a suitable target self-assembled monolayer (SAM) [31–32]. Amongst others, we reported carbohydrate microarrays fabricated by cycloaddition of alkynes on azide-terminated SAMs [33], by Diels–Alder reaction of cyclopentadienes and furans on maleimide-terminated SAMs [34], by thiol–ene click reaction of functionalized thiols on alkene-terminated SAMs [35] as well as by strain promoted cycloadditions on azide- and nitriloxide-terminated SAMs [36] using μCP. Homogenous spots, high-edge resolution, good reproducibility and short reaction times can be easily achieved by using µCP.…”

A method of printing uniform protein lines by using flat PDMS stamps

“…lCP is simple to apply and has good versatility [8,[19][20][21][22][23][24]. It was introduced by Kumar and coworkers in 1993 for replicating patterns generated by photolithography [25].…”

Patterned Fluorescence Images with Indigo Precursors in Polymer Film

“…Fabrication of fluorescence patterns is of great technological importance in a variety of fields including sensor, display and electronic industries. [1][2][3][4][5] Among the various techniques such as photoirradiation, [6][7][8][9] inkjet/screen printing, 10,11 vapor deposition, 12 and microcontact printing [13][14][15] methods developed to date, the photoirradiation method represents the most popular and practical approach for the fabrication of patterned fluorescence images on a solid support. In a typical photoirradiation method, functional molecules having photochemically removable protecting groups are immobilized or coated on the solid substrate.…”