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ACS Sens. 2017 Apr 28;2(4):495-500. doi: 10.1021/acssensors.7b00136. Epub 2017 Apr 17.

Steric Hindrance Assay for Secreted Factors in Stem Cell Culture.

Author information

1
Electrical and Computer Engineering, University of Toronto , Toronto M5S 1A4 Canada.
2
Leslie Dan Faculty of Pharmacy, University of Toronto , Toronto M5S 3M2 Canada.
3
Institute of Biomaterials and Biomedical Engineering, University of Toronto , Toronto M5S 3G9 Canada.
4
Départment de Chimie, Université de Montréal , Montreal H3T 1J4 Canada.
5
Department of Biochemistry, University of Toronto , Toronto M5S 1A8 Canada.

Abstract

The ex vivo expansion of hematopoietic stem cells is significantly inhibited by secreted proteins that induce negative feedback loops. The ability to effectively monitor these factors is critical for their real-time regulation and control and, by extension, enhancing stem cell expansion. Here, we describe a novel monitoring strategy for the detection of soluble signaling factors in stem cell cultures using a DNA-based sensing mechanism on a chip-based nanostructured microelectrode platform. We combine DNA hybridization engineering with antibody-capturing chemistry in an amplified steric hindrance hybridization assay. This method enables the quantification of important secreted proteins, showcased by the detection of 10 pg·mL-1 level concentrations of three proteins in stem cell culture samples. This approach is the first universal nonsandwich technique that permits pg·mL-1 level quantification of small proteins in stem cell culture media without signal loss.

KEYWORDS:

DNA; biosensors; electrochemistry; nanostructures; proteins; stem cells; steric hindrance

PMID:
28723184
DOI:
10.1021/acssensors.7b00136

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