Electrochemistry-based approaches to low cost, high sensitivity, automated, multiplexed protein immunoassays for cancer diagnostics

Dixit, Chandra Kumar; Kadimisetty, Karteek; Otieno, Brunah; Tang, Chi Kwong; Malla, Spundana; Krause, Colleen E.; Rusling, James F.

doi:10.1039/c5an01829c

Cited by 67 publications

(48 citation statements)

References 68 publications

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“…15–18 We have used antibodies in our laboratory in modern microfluidics devices to achieve ultra-sensitive multiplexed protein assay. 19–23 While a wide variety of antibodies are commercially available, isolation and purification makes them quite expensive, and storage stability can be an issue. 24 Recombinant technologies do not yet provide a general alternative.…”

Section: Introductionmentioning

confidence: 99%

Antibody-like Biorecognition Sites for Proteins from Surface Imprinting on Nanoparticles

Bhakta

Seraji

Suib

et al. 2015

ACS Appl. Mater. Interfaces

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Natural antibodies are used widely for important applications such as biomedical analysis, cancer therapy, and directed drug delivery, but they are expensive and may have limited stability. This study describes synthesis of antibody-like binding sites by molecular imprinting on silica nanoparticles (SiNP) using a combination of four organosilane monomers with amino acid-like side chains providing hydrophobic, hydrophilic, and H-bonding interactions with target proteins. This approach provided artificial antibody (AA) nanoparticles with good selectivity and specificity to binding domains on target proteins in a relatively low-cost synthesis. The AAs were made by polymer grafting onto SiNPs for human serum albumin (HSA) and glucose oxidase (GOx). Binding affinity, selectivity, and specificity was compared to several other proteins using adsorption isotherms and surface plasmon resonance (SPR). The Langmuir–Freundlich adsorption model was used to obtain apparent binding constants (KLF) from binding isotherms of HSA (6.7 × 104) and GOx (4.7 × 104) to their respective AAs. These values were 4–300 fold larger compared to a series of nontemplate proteins. SPR binding studies of AAs with proteins attached to a gold surface confirmed good specificity and revealed faster binding for the target proteins compared to nontarget proteins. Target proteins retained their secondary structures upon binding. Binding capacity of AAHSA for HSA was 5.9 mg HSA/g compared to 1.4 mg/g for previously report imprinted silica beads imprinted with poly(aminophenyl)boronic acid. Also, 90% recovery for HSA spiked into 2% calf serum was found for AAHSA.

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Section: Introductionmentioning

confidence: 99%

Antibody-like Biorecognition Sites for Proteins from Surface Imprinting on Nanoparticles

Bhakta

Seraji

Suib

et al. 2015

ACS Appl. Mater. Interfaces

Self Cite

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“…and evaluated as indicators of normal or abnormal processes or conditions or diseases and pharmacologic responses to therapeutics [4]. Concentration levels of these molecules are representative of changes occurring in the basic cell regulatory functions and cellular physiology, such as cell division or contact inhibition, and may vary from their normal levels in case of diseases and disorders [5]. Circulating biomarkers can derive from various molecular origins, including small molecules such as glucose or cholesterol, DNAs (i.e., specific mutation, translocation, amplification in characteristic genes such as BCRA1 , KRAS , cell-free DNAs, RNAs (mRNAs and miRNAs), proteins (i.e., hormones, antibodies, oncogenes, or tumor suppressors), exosomes and tumor cells.…”

Section: Circulating Biomarkersmentioning

confidence: 99%

Electrochemical Genosensing of Circulating Biomarkers

Campuzano

Yáñez‐Sedeño

Pingarrón

2017

Sensors

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Management and prognosis of diseases requires the measurement in non- or minimally invasively collected samples of specific circulating biomarkers, consisting of any measurable or observable factors in patients that indicate normal or disease-related biological processes or responses to therapy. Therefore, on-site, fast and accurate determination of these low abundance circulating biomarkers in scarcely treated body fluids is of great interest for health monitoring and biological applications. In this field, electrochemical DNA sensors (or genosensors) have demonstrated to be interesting alternatives to more complex conventional strategies. Currently, electrochemical genosensors are considered very promising analytical tools for this purpose due to their fast response, low cost, high sensitivity, compatibility with microfabrication technology and simple operation mode which makes them compatible with point-of-care (POC) testing. In this review, the relevance and current challenges of the determination of circulating biomarkers related to relevant diseases (cancer, bacterial and viral infections and neurodegenerative diseases) are briefly discussed. An overview of the electrochemical nucleic acid–based strategies developed in the last five years for this purpose is given to show to both familiar and non-expert readers the great potential of these methodologies for circulating biomarker determination. After highlighting the main features of the reported electrochemical genosensing strategies through the critical discussion of selected examples, a conclusions section points out the still existing challenges and future directions in this field.

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“…15 We recently utilized a SLA printer to fabricate a clear plastic flow sensor cell to measure DNA by electrochemiluminescence (ECL), 10 and 3D-printed a supercapacitor-powered electrochemiluminescent (ECL) immunoarray for multiplexed detection of 3 cancer biomarker proteins at fg mL −1 levels. 16 …”

Section: Introductionmentioning

confidence: 99%

Automated 3D-printed unibody immunoarray for chemiluminescence detection of cancer biomarker proteins

2017

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A low cost three-dimensional (3D) printed clear plastic microfluidic device was fabricated for fast, low cost automated protein detection. The unibody device features three reagent reservoirs, an efficient 3D network for passive mixing, and an optically transparent detection chamber housing a glass capture antibody array for measuring chemiluminescence output with a CCD camera. Sandwich type assays were built onto the glass arrays using a multi-labeled detection antibody-polyHRP (HRP = horseradish peroxidase). Total assay time was ~30 min in a complete automated assay employing a programmable syringe pump so that the protocol required minimal operator intervention. The device was used for multiplexed detection of prostate cancer biomarker proteins prostate specific antigen (PSA) and platelet factor 4 (PF-4). Detection limits of 0.5 pg mL−1 were achieved for these proteins in diluted serum with log dynamic ranges of four orders of magnitude. Good accuracy vs ELISA was validated by analyzing human serum samples. This prototype device holds good promise for further development as a point-of-care cancer diagnostics tool.

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Electrochemistry-based approaches to low cost, high sensitivity, automated, multiplexed protein immunoassays for cancer diagnostics

Cited by 67 publications

References 68 publications

Antibody-like Biorecognition Sites for Proteins from Surface Imprinting on Nanoparticles

Antibody-like Biorecognition Sites for Proteins from Surface Imprinting on Nanoparticles

Electrochemical Genosensing of Circulating Biomarkers

Automated 3D-printed unibody immunoarray for chemiluminescence detection of cancer biomarker proteins

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