Integration of Protein Processing Steps on a Droplet Microfluidics Platform for MALDI-MS Analysis

Chatterjee, Debalina; Son, Seok Man; Loo, Joseph A.; Garrell, Robin L.

doi:10.1021/ac9029373

Cited by 75 publications

(72 citation statements)

References 69 publications

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“…77 Here, we report an integrated method for implementing three discrete processing steps, including proteomic sample reduction, alkylation, and digestion; this paper joins related work 52,126,127 in demonstrating the compatibility of DMF with multistep proteomic sample processing. The qualitative and quantitative data presented here suggest that DMF is capable of highly repeatable sample processing, and thus may be a useful new tool for standardized proteomics analyses.…”

Section: Introductionmentioning

confidence: 80%

“…[49][50][51][52] In DMF, discrete microdroplets are manipulated electrostatically on an array of electrodes coated with a hydrophobic, dielectric insulator. 53,54 The classical characterization of droplet movement in DMF is based on a thermodynamic approach using the Young-Lippman equation (Eq 1), 55,56 where change in contact angle, , is a function of applied voltage, V. Here,   is the contact angle when the electric field across interfacial layer is zero, ε r is the relative permittivity of the dielectric, ε o is the permittivity of free space,  is the is the liquid-filler media surface tension, and d is the dielectric thickness.…”

Section: Digital Microfluidicsmentioning

confidence: 99%

“…DMF is a fluid-handling technique in which liquid samples are manipulated on the surface of an array of electrodes by electrostatic forces, 122 and has been applied to a variety of applications related to proteomics including sample purification by precipitation, 50 reduction of disulfides (chapter 3), 49,52 alkylation of thiols (chapter 3), 49,51,52 proteolytic digestion (chapters 2 and 3), 49,51,52,128 and combinations thereof. 163 These previous reports represent important steps towards the goal of integrated proteome profiling, but they are limited by the challenges inherent to homogeneous digestions described above.…”

Section: Introductionmentioning

confidence: 99%

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A Digital Microfluidic Approach to Proteomic Sample Processing

2009

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Proteome profiling is the identification and quantitation of all proteins in biological samples. An important application of proteome profiling that has received much attention is clinical proteomics, a field that promises the discovery of biomarkers that will be useful for early diagnosis and prognosis of diseases. While clinical proteomic methods vary widely, a common characteristic is the need for (i) extraction of proteins from complex biological fluids and (ii) extensive biochemical processing (reduction, alkylation and enzymatic digestion) prior to analysis. However, the lack of standardized sample handling and processing in proteomics is a major limitation for the field. The conventional macroscale manual sample handling requires multiple containers and transfers, which often leads to sample loss and contamination. For clinical proteomics to be adopted as a gold standard for clinical measures, the issue of irreproducibility needs to be addressed. A potential solution to this problem is to form integrated systems for sample handling and processing, and in this dissertation, I describe my work towards realizing this goal using digital microfluidics (DMF). DMF is a technique characterized by the manipulation of discrete droplets (100 nL -10 L) on an array of electrodes by the application of electrical fields. It is well-suited for carrying out rapid, sequential, miniaturized automated biochemical assays. This thesis demonstrates how DMF can be a powerful tool capable of automating several protein handling and processing steps used in proteomics.iii Methods for implementing proteomic multi-step solution-phase processes (reduction, alkylation, and digestion) on DMF were developed. This was accompanied by the development of heterogenous enzymatic microreactors for efficient protein digestion. Strategies for reducing non-specific adsorption were developed. Finally, in proof-of-concept work, a combination of protein extraction and protein processing was demonstrated on DMF devices.iv ACKNOWLEDGEMENTS

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Section: Introductionmentioning

confidence: 80%

Section: Digital Microfluidicsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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A Digital Microfluidic Approach to Proteomic Sample Processing

2009

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“…Similar to the digestion of lysozyme, the digestion of BSA by TY-microreactor at 50 °C efficiently occurred but not at 30 °C. The sequence coverage of 37% (201/583 amino acids) was higher compared with that of in-solution digestion (26%, 151/583 amino acids) and was better or comparable to those of a thermal (Liu et al, 2009;Sim et al, 2006) or chemical denatured BSA (Li et al, 2007a;Chatterjee et al, 2010) or the reduced BSA (Ma et al, 2008) by the reported trypsin-microreactors (24-46%). In addition, the number of identified disulfide bonds was 10 of 17, which is superior to 6 obtained by in-solution digestion.…”

Section: In Addition It I S P O S S I B L E T H a T O U R M S S Y S mentioning

confidence: 85%

Simple and Rapid Proteomic Analysis by Protease-Immobilized Microreactors

Yamaguchi¹,

Miyazaki²,

Maeda³

2012

Integrative Proteomics

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“…Chatterjee D et al showed that a complete integrated sequence of protein processing steps could be performed on droplet-based microfluidics platform, including disulfide reduction, alkylation, and enzymatic digestion, followed by cocrystallization with a MALDI matrix and analysis of the sample in situ by MALDI-MS [18]. In 2009, Jebrail M J et al reported the development of an automated microfluidic method for extracting proteins from heterogeneous fluids by precipitation [19].…”

Section: Peptide and Protein Analysismentioning

confidence: 99%

Biological Application of Digital Microfluidics Technology

Liu¹,

Deng²,

Qin³

et al. 2010

Nano BioMed ENG

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Digital microfluidics technology offers a platform for developing diagnostic applications with the advantages of portability, sample and reagent volume reduction, faster analysis, increased automation, low power consumption, compatibility with mass manufacturing and high throughput. In addition to diagnostics, digital microfluidics is finding use in nucleic acid analysis, peptide and protein analysis, cell analysis, drug analysis and delivery and immunization analysis. In this review, we describe these applications, their implementation, and associated design issues. As other review in the digital microfluidics technology, there have been and will be unexpected developments as DMF matures, but we predict that the future is bright for this promising technology at the last section.Keywords: Digital microfluidics technology; Nucleic acid analysis; Peptide and protein analysis; Cell analysis; Drug analysis and delivery Citation: B. Liu et al. Biological Application of Digital Microfluidics Technology. Nano Biomed Eng. 2010, 2(2), 149-154. DOI: 10.5101/nbe.v2i2.p149-154. The advent and development of digital microfluidics technologyTraditional (continuous-flow) microfluidic technologies are based on the continuous flow of liquid through microfabricated channels [1]. Continuous-flow systems are inherently difficult to integrate because the parameters that govern flow field (e.g. pressure, fluid resistance, electric field strength) vary along the flow-path, making the flow at any location dependent upon the properties of the entire system.The concept of digital microfluidics (DMF) arose in the late 1990s and involves the manipulation of discrete volumes of liquids on a surface. Manipulation of droplets can occur through various mechanisms, including electrowetting [2], dielectrophoresis [3], thermocapillary transport [4] and surface acoustic wave transport [5]. DMF was popularized in the early 2000s by Fair and coworkers [6] and Kim and coworkers [7] at Duke and UCLA, respectively. The technique was explained as being a phenomenon driven by surface tension, and was called "electrowetting" or "electrowetting ondielectric" (EWOD). A detailed review of electrowetting basics can be found in the work of Mugele [8]. In addition, work on simulation and modeling of dropletbased electrowetting has been reported by Biddut Bhattacharjee and Homayoun Najjaran [9].In contrast to continuous-flow biochips, digital microfluidic biochips platform is under software-driven electronic control, eliminating the need for mechanical tubes, pumps, and valves. Moreover, because each droplet can be controlled independently, these "digital" systems also have dynamic reconfigurability, whereby groups of cells in a microfluidic array can be reconfigured to change their functionality during the concur- Application of digital microfluidics technology in bioanalysisTo accomplish a digital microfluidics device it requires a hierarchical taxonomy. At the top level, applications are scaled to a microfluidic platform. The second level describ...

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Integration of Protein Processing Steps on a Droplet Microfluidics Platform for MALDI-MS Analysis

Cited by 75 publications

References 69 publications

A Digital Microfluidic Approach to Proteomic Sample Processing

A Digital Microfluidic Approach to Proteomic Sample Processing

Simple and Rapid Proteomic Analysis by Protease-Immobilized Microreactors

Biological Application of Digital Microfluidics Technology

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