Analytical and Bioanalytical Chemistry

, Volume 407, Issue 28, pp 8451–8462

Evaluating mixtures of 14 hygroscopic additives to improve antibody microarray performance

  • Sébastien Bergeron
  • Veronique Laforte
  • Pik-Shan Lo
  • Huiyan Li
  • David Juncker
Research Paper

DOI: 10.1007/s00216-015-8992-8

Cite this article as:
Bergeron, S., Laforte, V., Lo, PS. et al. Anal Bioanal Chem (2015) 407: 8451. doi:10.1007/s00216-015-8992-8

Abstract

Microarrays allow the miniaturization and multiplexing of biological assays while only requiring minute amounts of samples. As a consequence of the small volumes used for spotting and the assays, evaporation often deteriorates the quality, reproducibility of spots, and the overall assay performance. Glycerol is commonly added to antibody microarray printing buffers to decrease evaporation; however, it often decreases the binding of antibodies to the surface, thereby negatively affecting assay sensitivity. Here, combinations of 14 hygroscopic chemicals were used as additives to printing buffers for contact-printed antibody microarrays on four different surface chemistries. The ability of the additives to suppress evaporation was quantified by measuring the residual buffer volume in open quill pins over time. The seven best additives were then printed either individually or as a 1:1 mixture of two additives, and the homogeneity, intensity, and reproducibility of both the spotted protein and of a fluorescently labeled analyte in an assay were quantified. Among the 28 combinations on the four slides, many were found to outperform glycerol, and the best additive mixtures were further evaluated by changing the ratio of the two additives. We observed that the optimal additive mixture was dependent on the slide chemistry, and that it was possible to increase the binding of antibodies to the surface threefold compared to 50 % glycerol, while decreasing whole-slide coefficient of variation to 5.9 %. For the two best slides, improvements were made for both the limit of detection (1.6× and 5.9×, respectively) and the quantification range (1.2× and 2.1×, respectively). The additive mixtures identified here thus help improve assay reproducibility and performance, and might be beneficial to all types of microarrays that suffer from evaporation of the printing buffers.

Keywords

Antibody Protein Microarray Low evaporation Contact printing Hygroscopic Reproducibility 

Abbreviations

1,3-But

1,3-Butanediol

2,3-But

2,3-Butanediol

Ab

Antibody

ACM

Antibody colocalization microarray

AF

AlexaFluor

APTES

(3-Aminopropyl)triethoxysilane

cAb

Capture antibody

dAb

Detection antibody

DMSO

Dimethyl sulfoxide

DNA

Deoxyribonucleic acid

ELISA

Enzyme-linked immunosorbent assay

EtGly

Ethylene glycol

IgG

Immunoglobulin G

LOD

Limit of detection

OM

Orders of magnitude

PBS

Phosphate buffer saline

PBST

Phosphate buffer saline with 0.1 % Tween-20

PEG

Polyethylene glycol

PVA

Polyvinyl alcohol

Supplementary material

216_2015_8992_MOESM1_ESM.pdf (3.3 mb)
ESM 1(PDF 3366 kb)

Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  • Sébastien Bergeron
    • 1
    • 2
  • Veronique Laforte
    • 1
    • 2
    • 3
  • Pik-Shan Lo
    • 1
    • 2
  • Huiyan Li
    • 1
    • 2
  • David Juncker
    • 1
    • 2
    • 3
  1. 1.McGill University & Genome Quebec Innovation CentreMcGill UniversityMontrealCanada
  2. 2.Biomedical Engineering DepartmentMcGill UniversityMontrealCanada
  3. 3.Neurology and Neurosurgery DepartmentMcGill UniversityMontrealCanada

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