Multiplexed Electrochemical Cancer Diagnostics With Automated Microfluidics.

Microfluidic platforms can lead to miniaturisation, increased throughput and reduced reagent consumption, particularly when the processes are automated. Here, a programmable microcontroller is used for automation of a microfluidic platform configured to electrochemically determine the levels of 8 proteins simultaneously in complex liquid samples. The platform system is composed of a programmable Arduino microcontroller that controls inexpensive valve actuators, pump, magnetic stirrer and electronic display. The programmable microcontroller results in repeatable timing for each step in a complex assay protocol, such as sandwich immunoassays. Application of the platform is demonstrated using a multiplexed electrochemical immunoassay based on capture at the electrode surface of magnetic particles labelled with horseradish peroxidase and detection antibody. The multiplexed assay protocol is completed in less than 30 mins and results in detection of eight proteins associated with prostate cancer. The approach presented can be used to automate and simplify high-throughput screening campaigns, such as detection of multiple biomarkers in patient samples.

[1]  James F Rusling,et al.  Ultrasensitive immunosensor for cancer biomarker proteins using gold nanoparticle film electrodes and multienzyme-particle amplification. , 2009, ACS nano.

[2]  J. Rusling,et al.  On-line protein capture on magnetic beads for ultrasensitive microfluidic immunoassays of cancer biomarkers. , 2014, Biosensors & bioelectronics.

[3]  S. Hanash,et al.  Mining the plasma proteome for cancer biomarkers , 2008, Nature.

[4]  M. Stampfer,et al.  Insulin-like growth factor I (IGF-I), IGF-binding protein-3 and prostate cancer risk: epidemiological studies. , 2000, Growth hormone & IGF research : official journal of the Growth Hormone Research Society and the International IGF Research Society.

[5]  R. Zengerle,et al.  Microfluidic lab-on-a-chip platforms: requirements, characteristics and applications. , 2010, Chemical Society reviews.

[6]  W. Benedict,et al.  Pigment epithelium-derived factor: a potent inhibitor of angiogenesis. , 1999, Science.

[7]  D. Leech,et al.  Inexpensive Miniature Programmable Magnetic Stirrer from Reconfigured Computer Parts. , 2017 .

[8]  Albert Folch,et al.  3D-printed microfluidic automation. , 2015, Lab on a chip.

[9]  D. Leech,et al.  Cost-Effective Wireless Microcontroller for Internet Connectivity of Open-Source Chemical Devices , 2018, Journal of Chemical Education.

[10]  W. Isaacs,et al.  GOLPH2 and MYO6: Putative prostate cancer markers localized to the Golgi apparatus , 2008, The Prostate.

[11]  Damith E W Patabadige,et al.  Micro Total Analysis Systems: Fundamental Advances and Applications. , 2016, Analytical chemistry.

[12]  M. I. Hassan,et al.  Structural Model of Human PSA: A Target for Prostate Cancer Therapy , 2007, Chemical biology & drug design.

[13]  James F Rusling,et al.  Multiplexed electrochemical protein detection and translation to personalized cancer diagnostics. , 2013, Analytical chemistry.

[14]  Timothy D. Veenstra,et al.  Proteomic patterns: their potential for disease diagnosis , 2005, Molecular and Cellular Endocrinology.

[15]  G. Whitesides The origins and the future of microfluidics , 2006, Nature.

[16]  S. Varambally,et al.  Antibody-based detection of ERG rearrangement-positive prostate cancer. , 2010, Neoplasia.

[17]  James F. Rusling,et al.  Rapid Microfluidic Immunoassays of Cancer Biomarker Proteins Using Disposable Inkjet-Printed Gold Nanoparticle Arrays , 2013, ChemistryOpen.

[18]  Jinke Li,et al.  The prognostic value of vascular endothelial growth factor in ovarian cancer: a systematic review and meta-analysis. , 2013, Gynecologic oncology.

[19]  James F Rusling,et al.  Measurement of biomarker proteins for point-of-care early detection and monitoring of cancer. , 2010, The Analyst.

[20]  N. Morgan,et al.  Electrochemical immunosensors for detection of cancer protein biomarkers. , 2012, ACS nano.

[21]  J. Du,et al.  Association between CD14 Promoter -159C/T Polymorphism and the Risk of Sepsis and Mortality: A Systematic Review and Meta-Analysis , 2013, PloS one.

[22]  J. Gu,et al.  Prognostic role of serum AZGP1, PEDF and PRDX2 in colorectal cancer patients. , 2013, Carcinogenesis.

[23]  J. W. Findlay,et al.  Validation of immunoassays for bioanalysis: a pharmaceutical industry perspective. , 2000, Journal of pharmaceutical and biomedical analysis.

[24]  B. Lokeshwar,et al.  Insulin-like growth factors and their binding proteins in prostate cancer: cause or consequence? , 2006, Urologic oncology.

[25]  Andrew J. Vickers,et al.  Prostate-specific antigen and prostate cancer: prediction, detection and monitoring , 2008, Nature Reviews Cancer.

[26]  K. Bensalah,et al.  New circulating biomarkers for prostate cancer , 2008, Prostate Cancer and Prostatic Diseases.

[27]  James F Rusling,et al.  Ultrasensitive detection of cancer biomarkers in the clinic by use of a nanostructured microfluidic array. , 2012, Analytical chemistry.

[28]  C. Bonorino,et al.  CD14 Expression in the First 24h of Sepsis: Effect of −260C>T CD14 SNP , 2008, Immunological investigations.

[29]  Ali K Yetisen,et al.  Commercialization of microfluidic devices. , 2014, Trends in biotechnology.