Sample solution constraints on motor-driven diagnostic nanodevices

Korten, Slobodanka; Albet-Torres, Nuria; Paderi, Francesca; Siethoff, Lasse ten; Diez, Stefan; Korten, Till; Kronnie, Geertruy te; Månsson, Alf

doi:10.1039/c2lc41099k

Cited by 28 publications

(29 citation statements)

References 50 publications

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“…However, other motor systems, such as the kinesin-microtubule system, may also be used, enabling wide applicability due to different dependence of motor function on surface chemistry (cf. ref (36)) for the two systems. Furthermore, other detectors should be possible to study by similar approaches.…”

Section: New Methods For Evaluating Biosensor Performancementioning

confidence: 99%

Molecular Motor Propelled Filaments Reveal Light-Guiding in Nanowire Arrays for Enhanced Biosensing

et al. 2014

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Semiconductor nanowire arrays offer significant potential for biosensing applications with optical read-out due to their high surface area and due to the unique optical properties of one-dimensional materials. A challenge for optical read-out of analyte-binding to the nanowires is the need to efficiently collect and detect light from a three-dimensional volume. Here we show that light from fluorophores attached along several μm long vertical Al2O3 coated gallium phosphide nanowires couples into the wires, is guided along them and emitted at the tip. This enables effective collection of light emitted by fluorescent analytes located at different focal planes along the nanowire. We unequivocally demonstrate the light-guiding effect using a novel method whereby the changes in emitted fluorescence intensity are observed when fluorescent cytoskeletal filaments are propelled by molecular motors along the wires. The findings are discussed in relation to nanobiosensor developments, other nanotechnological applications, and fundamental studies of motor function.

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Section: New Methods For Evaluating Biosensor Performancementioning

confidence: 99%

Molecular Motor Propelled Filaments Reveal Light-Guiding in Nanowire Arrays for Enhanced Biosensing

et al. 2014

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“…This is a concern as we recently [22] found that both actomyosin and microtubule-kinesin sliding velocity is effectively reduced or inhibited by body fluids (blood components, cell lysates) unless diluted 100–200 times. This finding is expanded on in Figure 1 where we, in addition to effects on velocity, also show concentration-dependent effects of serum on the fraction of motile filaments and on the fraction of independent experiments where actomyosin motility was observed.…”

Section: Resultsmentioning

confidence: 99%

“…Different factors are likely to contribute to the deleterious effects. In addition to serum proteins such as gelsolin, directly depolymerising the actin filaments [27], there may be binding of other blood serum components to HMM or actin [22]. …”

Section: Resultsmentioning

confidence: 99%

“…Particularly, we showed recently [22] that complex fluid environments, such as blood plasma, blood serum and cell lysates may have deleterious effects on molecular motor driven propulsion of both actin filaments and microtubules, unless the samples are diluted > 100 times. These problems are essential to overcome since extensive sample dilution is highly undesirable in high-sensitivity detection.…”

Section: Introductionmentioning

confidence: 99%

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Magnetic capture from blood rescues molecular motor function in diagnostic nanodevices

et al. 2013

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BackgroundIntroduction of effective point-of-care devices for use in medical diagnostics is part of strategies to combat accelerating health-care costs. Molecular motor driven nanodevices have unique potentials in this regard due to unprecedented level of miniaturization and independence of external pumps. However motor function has been found to be inhibited by body fluids.ResultsWe report here that a unique procedure, combining separation steps that rely on antibody-antigen interactions, magnetic forces applied to magnetic nanoparticles (MPs) and the specificity of the actomyosin bond, can circumvent the deleterious effects of body fluids (e.g. blood serum). The procedure encompasses the following steps: (i) capture of analyte molecules from serum by MP-antibody conjugates, (ii) pelleting of MP-antibody-analyte complexes, using a magnetic field, followed by exchange of serum for optimized biological buffer, (iii) mixing of MP-antibody-analyte complexes with actin filaments conjugated with same polyclonal antibodies as the magnetic nanoparticles. This causes complex formation: MP-antibody-analyte-antibody-actin, and magnetic separation is used to enrich the complexes. Finally (iv) the complexes are introduced into a nanodevice for specific binding via actin filaments to surface adsorbed molecular motors (heavy meromyosin). The number of actin filaments bound to the motors in the latter step was significantly increased above the control value if protein analyte (50–60 nM) was present in serum (in step i) suggesting appreciable formation and enrichment of the MP-antibody-analyte-antibody-actin complexes. Furthermore, addition of ATP demonstrated maintained heavy meromyosin driven propulsion of actin filaments showing that the serum induced inhibition was alleviated. Detailed analysis of the procedure i-iv, using fluorescence microscopy and spectroscopy identified main targets for future optimization.ConclusionThe results demonstrate a promising approach for capturing analytes from serum for subsequent motor driven separation/detection. Indeed, the observed increase in actin filament number, in itself, signals the presence of analyte at clinically relevant nM concentration without the need for further motor driven concentration. Our analysis suggests that exchange of polyclonal for monoclonal antibodies would be a critical improvement, opening for a first clinically useful molecular motor driven lab-on-a-chip device.

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“…Hence, a detection of microvesicles in blood, serum, saliva, and other biological fluids, and analysis of microvesiclecarried cargo pattern represents a novel, noninvasive diagnostic approach (2,(20)(21)(22). In this regard, lab-on-a-chip systems represent a promising and intriguing method using molecular motor-driven diagnostics for sensitive detection of biomarkers in extremely small sample volumes (23). We recently investigated high-capacity nanocarriers, such as actin filaments or bundles, for microvesicle detection (24).…”

Section: Introductionmentioning

confidence: 99%

Expression Profiling of Circulating Microvesicles Reveals Intercellular Transmission of Oncogenic Pathways

Milani

Lana

Bresolin

et al. 2017

Molecular Cancer Research

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Circulating microvesicles have been described as important players in cell-to-cell communication carrying biological information under normal or pathologic condition. Microvesicles released by cancer cells may incorporate diverse biomolecules (e.g., active lipids, proteins, and RNA), which can be delivered and internalized by recipient cells, potentially altering the gene expression of recipient cells and eventually impacting disease progression. Leukemia in vitro model systems were used to investigate microvesicles as vehicles of protein-coding messages. Several leukemic cells (K562, LAMA-87, TOM-1, REH, and SHI-1), each carrying a specific chromosomal translocation, were analyzed. In the leukemic cells, these chromosomal translocations are transcribed into oncogenic fusion transcripts and the transfer of these transcripts was monitored from leukemic cells to microvesicles for each of the cell lines. Microarray gene expression profiling was performed to compare transcriptomes of K562-derived microvesicles and parental K562 cells. The data show that oncogenic BCR-ABL1 transcripts and mRNAs related to basic functions of leukemic cells were included in microvesicles. Further analysis of microvesicles cargo revealed a remarkable enrichment of transcripts related to cell membrane activity, cell surface receptors, and extracellular communication when compared with parental K562 cells. Finally, coculturing of healthy mesenchymal stem cells (MSC) with K562-derived microvesicles displayed the transfer of the oncogenic message, and confirmed the increase of target cell proliferation as a function of microvesicle dosage.Implications: This study provides novel insight into tumorderived microvesicles as carriers of oncogenic protein-coding messages that can potentially jeopardize cell-directed therapy, and spread to other compartments of the body. Mol Cancer Res; 15(6); 683-95. Ó2017 AACR.

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Sample solution constraints on motor-driven diagnostic nanodevices

Cited by 28 publications

References 50 publications

Molecular Motor Propelled Filaments Reveal Light-Guiding in Nanowire Arrays for Enhanced Biosensing

Molecular Motor Propelled Filaments Reveal Light-Guiding in Nanowire Arrays for Enhanced Biosensing

Magnetic capture from blood rescues molecular motor function in diagnostic nanodevices

Expression Profiling of Circulating Microvesicles Reveals Intercellular Transmission of Oncogenic Pathways

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