Antibody Colocalization Microarray for Cross-Reactivity-Free Multiplexed Protein Analysis

  • Véronique Laforte
  • Pik-Shan Lo
  • Huiyan Li
  • David JunckerEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1619)


Measuring many proteins at once is of great importance to the idea of personalized medicine, in order to get a snapshot of a person’s health status. We describe the antibody colocalization microarray (ACM), a variant of antibody microarrays which avoids reagent-induced cross-reactivity by printing individual detection antibodies atop their corresponding capture antibodies. We discuss experimental parameters that are critical for the success of ACM experiments, namely, the printing positional accuracy needed for the two printing rounds and the need for protecting dried spots during the second printing round. Using small sample volumes (less than 30 μL) and small quantities of reagents, up to 108 different targets can be measured in hundreds of samples with great specificity and sensitivity.

Key words

Microarray Antibody Sandwich immunoassay Multiplexed Fluorescence 



We wish to thank the National Science and Engineering Research Council (NSERC) of Canada, the Canadian Cancer Society (CCS), and the Canadian Institute of Health Research (CIHR) for funding. D.J. holds a Canada Research Chair (CRC) in Micro- and Nanobioengineering.


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Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  • Véronique Laforte
    • 1
    • 2
    • 3
  • Pik-Shan Lo
    • 2
    • 3
  • Huiyan Li
    • 2
    • 3
  • David Juncker
    • 1
    • 2
    • 3
    • 4
    Email author
  1. 1.Department of Neurology and Neurosurgery, Montreal Neurological InstituteMcGill UniversityMontrealCanada
  2. 2.Department of Biomedical EngineeringMcGill UniversityMontrealCanada
  3. 3.McGill University and Genome Quebec Innovation CenterMcGill UniversityMontrealCanada
  4. 4.Micro and Nanobioengineering Laboratory, Department of Biomedical EngineeringMcGill UniversityMontrealCanada

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