ArticleMolecular & cellular proteomics : MCP2012
Antibody colocalization microarray: a scalable technology for multiplex protein analysis in complex samples.
Article in Molecular & cellular proteomics : MCP, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 34 papers.
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Who cites it
34 citing papers in PubMed, 84 citations in OpenAlex.
- Expanding High-Fidelity Multiplexing in Ultrasensitive Single-Molecule Protein Detection via Proximity Barcoding.Angewandte Chemie (International ed. in English) · 2026Article
- Kinetic Profiling in One-Step Digital Immunoassays Enables Multiplex Quantification across an Ultrabroad Dynamic Range.Journal of the American Chemical Society · 2026Article
- nELISA: a high-throughput, high-plex platform enables quantitative profiling of the inflammatory secretome.Nature methods · 2025Article
- Evaluating Cross-Reactivity in Custom-Made Multiplex Bead-Based Antibody Microarrays: A Methodological Approach.Methods in molecular biology (Clifton, N.J.) · 2025Article
- A Multiplexed Digital Platform Enables Detection of Attomolar Protein Levels with Minimal Cross-Reactivity.ACS nano · 2024Article
- Article
- Generation of single stranded DNA with selective affinity to bovine spermatozoa.Animal bioscience · 2021Article
- Sequential Protein Capture in Multiplex Single Molecule Arrays: A Strategy for Eliminating Assay Cross-Reactivity.Advanced healthcare materials · 2021Article
- Composable Microfluidic Plates (cPlate): A Simple and Scalable Fluid Manipulation System for Multiplexed Enzyme-Linked Immunosorbent Assay (ELISA).Analytical chemistry · 2021Article
- The acute respiratory distress syndrome biomarker pipeline: crippling gaps between discovery and clinical utility.Translational research : the journal of laboratory and clinical medicine · 2020Review
- Multiplexed Immunosensor Based on the Amperometric Transduction for Monitoring of Marine Pollutants in Sea Water.Sensors (Basel, Switzerland) · 2020Article
- Article
- Comparison of Bead-Based Fluorescence Versus Planar Electrochemiluminescence Multiplex Immunoassays for Measuring Cytokines in Human Plasma.Frontiers in immunology · 2020Article
- A self-assembling peptide hydrogel for ultrarapid 3D bioassays.Nanoscale advances · 2019Article
- Ensemble multicolour FRET model enables barcoding at extreme FRET levels.Nature nanotechnology · 2018Article
- Current applications of antibody microarrays.Clinical proteomics · 2018Review
- Snap Chip for Cross-reactivity-free and Spotter-free Multiplexed Sandwich Immunoassays.Journal of visualized experiments : JoVE · 2017Article
- Prediction of Response to Sorafenib in Hepatocellular Carcinoma: A Putative Marker Panel by Multiple Reaction Monitoring-Mass Spectrometry (MRM-MS).Molecular & cellular proteomics : MCP · 2017Article
- Three-Dimensionally Functionalized Reverse Phase Glycoprotein Array for Cancer Biomarker Discovery and Validation.Journal of the American Chemical Society · 2016Article
- Bulked sample analysis in genetics, genomics and crop improvement.Plant biotechnology journal · 2016Review
Corrections and comments
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Authors and funding
11 authors at 3 institutions in 1 country.
Funding
Abstract
DNA microarrays were rapidly scaled up from 256 to 6.5 million targets, and although antibody microarrays were proposed earlier, sensitive multiplex sandwich assays have only been scaled up to a few tens of targets. Cross-reactivity, arising because detection antibodies are mixed, is a known weakness of multiplex sandwich assays that is mitigated by lengthy optimization. Here, we introduce (1) vulnerability as a metric for assays. The vulnerability of multiplex sandwich assays to cross-reactivity increases quadratically with the number of targets, and together with experimental results, substantiates that scaling up of multiplex sandwich assays is unfeasible. We propose (2) a novel concept for multiplexing without mixing named antibody colocalization microarray (ACM). In ACMs, both capture and detection antibodies are physically colocalized by spotting to the same two-dimensional coordinate. Following spotting of the capture antibodies, the chip is removed from the arrayer, incubated with the sample, placed back onto the arrayer and then spotted with the detection antibodies. ACMs with up to 50 targets were produced, along with a binding curve for each protein. The ACM was validated by comparing it to ELISA and to a small-scale, conventional multiplex sandwich assay (MSA). Using ACMs, proteins in the serum of breast cancer patients and healthy controls were quantified, and six candidate biomarkers identified. Our results indicate that ACMs are sensitive, robust, and scalable.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.