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Int J Nanomedicine. 2015 Nov 24;10:7197-205. doi: 10.2147/IJN.S92596. eCollection 2015.

Enhanced detection of single-cell-secreted proteins using a fluorescent immunoassay on the protein-G-terminated glass substrate.

Author information

1
Department of Bionano Technology, Graduate School, Hanyang University, Seoul, South Korea ; Department of Bionano Engineering, Hanyang University ERICA, Ansan, South Korea.
2
School of Integrative Engineering, Chung-Ang University, Seoul, South Korea.

Abstract

We present an evaluation of protein-G-terminated glass slides that may contain a suitable substrate for aligning the orientation of antibodies to obtain better binding moiety to the target antigen. The results of the protein-G-terminated slides were compared with those obtained with epoxy-based slides to evaluate signal enhancement for human immunoglobulin G (IgG) targets, and an increase in the average fluorescence intensity was observed for the lowest measurable amount of IgG target in the assay using protein-G-terminated slides. Applying this strategy for signal amplification to single-cell assays improves the limits of detection for human IgG protein and cytokines (interleukin-2 and interferon-γ) captured from hybridomas. Our data indicate that protein-G-terminated slides have a higher binding capacity for antigens and have better spot-to-spot consistency than that of traditional epoxy-based slides. These properties would be beneficial in the detection of fine amounts of single-cell-secreted proteins, which may provide key insights into cell-cell communication and immune responses.

KEYWORDS:

antibody’s orientation; microwell array; protein-G-terminated surface; secreted cytokine; single cell analysis

PMID:
26648723
PMCID:
PMC4664541
DOI:
10.2147/IJN.S92596
[Indexed for MEDLINE]
Free PMC Article

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