Patterning Multiplex Protein Microarrays in a Single Microfluidic Channel

• Published in: Analytical chemistry (Washington) Vol. 84; no. 2; pp. 1012 - 1018
• Main Authors: Didar, Tohid Fatanat, Foudeh, Amir M, Tabrizian, Maryam
• Format: Journal Article
• Language: English
• Published: Washington, DC American Chemical Society 17.01.2012
• Subjects: Analytical chemistry; antibodies; bioassays; biochemical compounds; Biological and medical sciences; Biosensors; Biotechnology; Chemistry; Exact sciences and technology; Flow rates; Fluorescence; Fluorescence microscopy; Fundamental and applied biological sciences. Psychology; General, instrumentation; High flow; Immunoassay; immunoassays; Methods. Procedures. Technologies; microarray technology; Microfluidic Analytical Techniques - instrumentation; Microfluidics; Microscopy, Fluorescence; Miscellaneous; Photoelectron Spectroscopy; Protein Array Analysis; Proteins; Proteins - chemistry; Shear stress; Spectrometric and optical methods; Spectrum analysis; Surface Properties; Various methods and equipments; X-ray photoelectron spectroscopy; Chemical analysis; Choice; Cross section; Microanalysis; X ray; Immunological method; Functionalization; Lead; Photoelectron spectrometry; Coupling; Printing; Fluid mechanics; Antibody; Device; X ray spectrometry; System on a chip; Chemical sensor; Protein; Experimental design; Biosensor; Sensor array; Application; Interface; Microfluidics; Fluorescence microscopy
• ISSN: 0003-2700; 1520-6882; 1520-6882
• DOI: 10.1021/ac2025877

The development of versatile biofunctional surfaces is a fundamental prerequisite in designing Lab on a Chip (LOC) devices for applications in biosensing interfaces and microbioreactors. The current paper presents a rapid combinatorial approach to create multiplex protein patterns in a single microfluidic channel. This approach consists of coupling microcontact printing with microfluidic patterning, where microcontact printing is employed for silanization using (3-Aminopropyl) triethoxysilane (APTES), followed by microfluidic patterning of multiple antibodies. As a result, the biomolecules of choice could be covalently attached to the microchannel surface, thus creating a durable and highly resistant functional interface. Moreover, the experimental procedure was designed to create a microfluidic platform that maintains functionality at high flow rates. The functionalized surfaces were characterized using X-ray photoelectron spectroscopy (XPS) and monitored with fluorescence microscopy at each step of functionalization. To illustrate the possibility of patterning multiple biomolecules along the cross section of a single microfluidic channel, microarrays of five different primary antibodies were patterned onto a single channel and their functionality was evaluated accordingly through a multiplex immunoassay using secondary antibodies specific to each patterned primary antibody. The resulting patterns remained stable at shear stresses of up to 50 dyn/cm2. The overall findings suggest that the developed multiplex functional interface on a single channel can successfully lead to highly resistant multiplex functional surfaces for high throughput biological assays.