Implementation of a Multiplex and Quantitative Proteomics Platform for Assessing Protein Lysates Using DNA-Barcoded Antibodies
| Autor: | Joseph M. Beechem, Yiling Lu, Huifang Guo, Lisa Y. Bogatzki, Rhonda Meredith, Savitri Krishnamurthy, Jinho Lee, Gordon B. Mills, Christopher P. Vellano, Shuangxing Yu, Gary K. Geiss, Zhiyong Ding, Warren Carter, Gokhan Demirkan, Zhenlin Ju, Brian Filanoski |
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| Rok vydání: | 2018 |
| Předmět: |
Proteomics
0301 basic medicine medicine.medical_treatment Systems biology Quantitative proteomics Computational biology Biology Biochemistry Antibodies Analytical Chemistry Targeted therapy 03 medical and health sciences Cell Line Tumor Neoplasms medicine Humans Multiplex Molecular Biology business.industry Technological Innovation and Resources Proteins Cancer DNA medicine.disease 030104 developmental biology Protein microarray Female Personalized medicine business |
| Zdroj: | Molecular & Cellular Proteomics. 17:1245-1258 |
| ISSN: | 1535-9476 |
| DOI: | 10.1074/mcp.ra117.000291 |
| Popis: | Molecular analysis of tumors forms the basis for personalized cancer medicine and increasingly guides patient selection for targeted therapy. Future opportunities for personalized medicine are highlighted by the measurement of protein expression levels via immunohistochemistry, protein arrays, and other approaches; however, sample type, sample quantity, batch effects, and "time to result" are limiting factors for clinical application. Here, we present a development pipeline for a novel multiplexed DNA-labeled antibody platform which digitally quantifies protein expression from lysate samples. We implemented a rigorous validation process for each antibody and show that the platform is amenable to multiple protocols covering nitrocellulose and plate-based methods. Results are highly reproducible across technical and biological replicates, and there are no observed "batch effects" which are common for most multiplex molecular assays. Tests from basal and perturbed cancer cell lines indicate that this platform is comparable to orthogonal proteomic assays such as Reverse-Phase Protein Array, and applicable to measuring the pharmacodynamic effects of clinically-relevant cancer therapeutics. Furthermore, we demonstrate the potential clinical utility of the platform with protein profiling from breast cancer patient samples to identify molecular subtypes. Together, these findings highlight the potential of this platform for enhancing our understanding of cancer biology in a clinical translation setting. |
| Databáze: | OpenAIRE |
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