Hydrogel filled silicon stamps for generating multiplexed protein microarrays

Date

2011-11-28

Author

Bat, Erhan
Huskens, Jurriaan

Metadata

Show full item record

Item Usage Stats

77
views

0
downloads

Protein microarrays have recently gained great interest as they are suited for performing bioanalytical applications such as in vitro diagnostics, proteomics, antibody characterization and drug screening. Several techniques such as ink-jet printing, dip-pen nanolithography and microcontact printing have been used to generate protein microarrays. Inkjet printing and dip pen nanolithography allow printing of multiple proteins however these are serial techniques, control over the spot geometry/size and reproducibility is challenging. Moreover the most commonly applied technique of inkjet printing does not allow printing of proteins with less than 50 µm resolution. Microcontact printing is a soft lithography technique that allows parallel arraying of a single biomolecule over a large surface area with high resolution. Microarrays with smaller feature sizes are desired as they would allow rapid, highly sensitive, and direct determination of analyte concentration using a small amount of sample. Microcontact printing is a low cost, simple and reproducible method but usually re-inking of the stamp is required in each replication step, which is time consuming and costly and creating a multiplexed array is still a challenge. In this project, we aim at addressing the multiplexicity and inking challenges of microcontact printing. We present hydrogel-filled silicon stamps having individually addressable ink reservoirs. With this design, multiplexicity can be achieved while avoiding the need for re-inking. To prepare the stamps, we fabricated silicon microstructures having separate wells (320×320×380 µm) and each well having a 25 µm thick membrane (144 microchannels measuring 5 µm in diameter) on the printing side. The reservoirs and the microchannels of the silicon microstructures were filled with macroporous poly(2-hydroxyethyl methacrylate-co-ethylene glycol dimethacrylate) hydrogels. Methacryloxypropyl silane functionalized wells allowed covalent binding of hydrogels to the surface of the silicon. Using these hydrogel-filled stamps, we printed arrays of fluorescently labeled immunoglobulins on polydimethylsiloxane substrates reproducibly at least up to twenty times without re-inking. The spot sizes were very close to 5 µm in agreement with the stamp design. The fluorescence intensities of the printed spots were comparable for each printing step indicating that the amount of transferred protein did not vary significantly. With the help of an inkjet spotter, we addressed three immunoglobulins each labelled with a different fluorophore to separate reservoirs of the stamp. In this manner, multiplexed protein microarrays were obtained. We have also evaluated this printing system in an antibody-antigen assay. These hydrogel-filled silicon stamps hold great promise for multiplexed bioanalytical applications as well as for patterning of bioactive molecules for tissue engineering applications.

URI

https://hdl.handle.net/11511/87003
https://www.mrs.org/fall2011/symposium-sessions/?code=II

Conference Name

2011 MRS Fall Meeting & Exhibit, November 28-December 2 2011

Collections

Department of Chemical Engineering, Conference / Seminar

Suggestions

OpenMETU
Core

Microfibrous scaffolds from poly(L-lactide-co-epsilon-caprolactone) blended with xeno-free collagen/hyaluronic acid for improvement of vascularization in tissue engineering applications KENAR, HALİME; Ozdogan, Candan Yilmaz; Dumlu, Cansu; DOĞER, EMEK; Kose, Gamze Torun; Hasırcı, Vasıf Nejat (2019-04-01) Success of 3D tissue substitutes in clinical applications depends on the presence of vascular networks in their structure. Accordingly, research in tissue engineering is focused on the stimulation of angiogenesis or generation of a vascular network in the scaffolds prior to implantation. A novel, xeno-free, collagen/hyaluronic acid-based poly(L-lactide-co-epsilon-caprolactone) (PLC/COL/HA) (20/9.5/0.5 w/w/w) microfibrous scaffold was produced by electrospinning. Collagen types I and III, and hyaluronic acid...
Multi-view subcellular localization prediction of human proteins Özsarı, Gökhan; Atalay, M. Volkan.; Department of Computer Engineering (2019) Determining the subcellular localization of proteins is crucial for Understanding the functions of proteins, drug targeting, systems biology, and proteomics research. Experimental validation of subcellular localization is an expensive and challenging process. There exist several computational methods for automated prediction of protein subcellular localization; however, there is still room for better performance. Here, we propose a multi-view SVM-based approach that provides predictions for human proteins. ...
Capillary electrophoresis with online stacking in combination with AgNPs@MCM-41 reinforced hollow fiber solid-liquid phase microextraction for quantitative analysis of Capecitabine and its main metabolite 5-Fluorouracil in plasma samples isolated from cancer patients Forough, Mehrdad; Farhadi, Khalil; Molaei, Rahim; Khalili, Hedayat; Shakeri, Ramin; Zamani, Asghar; Matin, Amir Abbas (2017-01-01) The purpose of this study is the development and validation of a simple, novel, selective and fast off-line microextraction technique combining capillary electrophoresis with in-column field-amplified sample injection (FASI) for the simultaneous determination of capecitabine (CAP) and its active metabolite, 5-Fluorouracil (5-FU), in human plasma. At the moment, there is a lack of using cost-effective CE tool combined with novel miniaturized sample clean-up techniques for analysis of these important anticanc...
Inference of Gene Regulatory Networks Via Multiple Data Sources and a Recommendation Method Ozsoy, Makbule Gulcin; Polat, Faruk; Alhajj, Reda (2015-11-12) Gene regulatory networks (GRNs) are composed of biological components, including genes, proteins and metabolites, and their interactions. In general, computational methods are used to infer the connections among these components. However, computational methods should take into account the general features of the GRNs, which are sparseness, scale-free topology, modularity and structure of the inferred networks. In this work, observing the common aspects between recommendation systems and GRNs, we decided to ...
Highly sensitive and selective colorimetric probe for determination of L-cysteine in aqueous media based on Ag/Pd bimetallic nanoparticles Farhadi, Khalil; Forough, Mehrdad; Pourhossein, Atefeh; Molaei, Rahim (2014-10-01) The interaction between Ag/Pd bimetallic nanoparticles and cysteine was introduced as a new and high potential colorimetric probe for the selective, sensitive and low-cost recognition and monitoring of cysteine in aqueous samples. In the presence of NaCl, cysteine was able to induce the aggregation of Ag/Pd bimetallic nanoparticles thereby resulting in a change in yellowish-brown color of the Ag/Pd colloid to green. The presence of 6.6 mM NaCl in the samples decreases the electrostatic repulsion and acceler...

Citation Formats

E. Bat and J. Huskens, “Hydrogel filled silicon stamps for generating multiplexed protein microarrays,” presented at the 2011 MRS Fall Meeting & Exhibit, November 28-December 2 2011, Boston, MA, USA, 2011, Accessed: 00, 2021. [Online]. Available: https://hdl.handle.net/11511/87003.