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

Autor: Laforte V; Department of Neurology and Neurosurgery, Montreal Neurological Institute, McGill University, 3801 University St., Montreal, QC, Canada, H3A 2B4.; Department of Biomedical Engineering, McGill University, 3775 University St., Montreal, QC, Canada, H3A 2B4.; McGill University and Genome Quebec Innovation Center, McGill University, 740 Dr. Penfield Avenue, Montreal, QC, Canada, H3A 0G1., Lo PS; Department of Biomedical Engineering, McGill University, 3775 University St., Montreal, QC, Canada, H3A 2B4.; McGill University and Genome Quebec Innovation Center, McGill University, 740 Dr. Penfield Avenue, Montreal, QC, Canada, H3A 0G1., Li H; Department of Biomedical Engineering, McGill University, 3775 University St., Montreal, QC, Canada, H3A 2B4.; McGill University and Genome Quebec Innovation Center, McGill University, 740 Dr. Penfield Avenue, Montreal, QC, Canada, H3A 0G1., Juncker D; Department of Neurology and Neurosurgery, Montreal Neurological Institute, McGill University, 3801 University St., Montreal, QC, Canada, H3A 2B4. david.juncker@mcgill.ca.; Department of Biomedical Engineering, McGill University, 3775 University St., Montreal, QC, Canada, H3A 2B4. david.juncker@mcgill.ca.; McGill University and Genome Quebec Innovation Center, McGill University, 740 Dr. Penfield Avenue, Montreal, QC, Canada, H3A 0G1. david.juncker@mcgill.ca.; Micro and Nanobioengineering Laboratory, Department of Biomedical Engineering, McGill University, 740 Dr. Penfield #6206, Montreal, QC, Canada, H3A 0G1. david.juncker@mcgill.ca.
Jazyk: angličtina
Zdroj: Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2017; Vol. 1619, pp. 239-261.
DOI: 10.1007/978-1-4939-7057-5_19
Abstrakt: 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.
Databáze: MEDLINE
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