Double chip protein arrays using recombinant single‐chain Fv antibody fragments

Gilbert, Ilka; Schiffmann, Susanne; Rubenwolf, Susanne; Jensen, Kristian; Mai, Thao; Albrecht, Christian; Lankenau, Andreas; Beste, Gerald; Blank, Kerstin; Gaub, Hermann E.; Clausen‐Schaumann, Hauke

doi:10.1002/pmic.200300736

Cited by 15 publications

(12 citation statements)

References 13 publications

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“…However, the logistic issues involved with scaling up such platforms combined with the observation that a threshold of about 50 probes per sandwich array might be the upper limit to still maintain adequate assay features (Haab, 2003), makes it a less attractive option when designing high-density microarrays. However, a novel array design, the double-chip antibody (protein) arrays-force-based multiplex sandwich assays-may provide one solution to this problem (Blank et al, 2004;Gilbert et al, 2003Gilbert et al, , 2004. In this set-up, a second chip surface is used for the local application of detection antibodies, thereby efficiently reducing any potential antibody cross-reactivities.…”

Section: Recent Advancesmentioning

confidence: 99%

Antibody Microarrays: Current Status and Key Technological Advances

Wingren

Borrebaeck

2006

OMICS: A Journal of Integrative Biology

101

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Antibody-based microarrays are among the novel classes of rapidly evolving proteomic technologies that holds great promise in biomedicine. Miniaturized microarrays (< 1 cm2) can be printed with thousands of individual antibodies carrying the desired specificities, and with biological sample (e.g., an entire proteome) added, virtually any specifically bound analytes can be detected. While consuming only minute amounts (< microL scale) of reagents, ultra- sensitive assays (zeptomol range) can readily be performed in a highly multiplexed manner. The microarray patterns generated can then be transformed into proteomic maps, or detailed molecular fingerprints, revealing the composition of the proteome. Thus, protein expression profiling and global proteome analysis using this tool will offer new opportunities for drug target and biomarker discovery, disease diagnostics, and insights into disease biology. Adopting the antibody microarray technology platform, several biomedical applications, ranging from focused assays to proteome-scale analysis will be rapidly emerging in the coming years. This review will discuss the current status of the antibody microarray technology focusing on recent technological advances and key issues in the process of evolving the methodology into a high-performing proteomic research tool.

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Section: Recent Advancesmentioning

confidence: 99%

Antibody Microarrays: Current Status and Key Technological Advances

Wingren

Borrebaeck

2006

OMICS: A Journal of Integrative Biology

101

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“…One of the goals of the HUPO Plasma Proteome Project (PPP), which was piloted in 2002, is to analyze comprehensively the protein constituents of human plasma and serum [12]. As a result of this initiative, several of the challenges associated with studying plasma, serum, and their constituents, such as lipoproteins, have been highlighted.…”

Section: Introductionmentioning

confidence: 99%

Quantitative comparison of lipoprotein fractions derived from human plasma and serum by liquid chromatography-tandem mass spectrometry

Collins

Olivier

2010

Proteome Sci

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BackgroundLipoproteins are complex, globular molecules which play essential roles in the transport and metabolism of cholesterol. Their implication in the development of cardiovascular diseases, such as atherosclerosis, has sustained a great deal of interest in these particles. Their various functions, and their contribution to the development of atherosclerosis, are often attributed to their protein constituents, which vary greatly among the different lipoprotein classes. Recent advances in the field of mass spectrometry have provided insight into the array of proteins associated with low-density lipoproteins (LDLs) and, even more so, with high-density lipoproteins (HDLs). Plasma and serum are the most commonly used samples for the analysis of lipoproteins. Although these lipoprotein sources are unique, it was our goal to determine whether or not their inherent differences would ultimately affect a quantitative analysis of the LDL and HDL proteomes. To this end, we isolated LDL and HDL fractions with fast protein liquid chromatography-size exclusion chromatography (FPLC-SEC) from both human plasma and serum samples from the same individuals. After delipidating these samples, we performed a quantitative proteomic analysis to compare the lipoprotein-associated proteins derived from both plasma and serum.ResultsAlthough the primary differences between the samples are found in fibrinogen proteins which are removed from serum, it of interest to note that, with respect to LDL-associated proteins, apolipoproteinB-100 was found at significantly higher levels in serum samples. Complement component 3 was found at significantly higher levels in serum-derived HDL fractions. Both of these proteins are known LDL- and HDL-associated proteins, respectively.ConclusionOverall, the results from our study indicate that both plasma and serum samples are equally suited for proteomic studies, and provide similar results. These findings are particularly important for studies profiling proteomic differences in lipoprotein particle composition in a variety of disease conditions, including cardiovascular disease.

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“…DNA‐based MFSs have not only been used for the detection of a number of different DNA modifications and interactions. The parallelization capacity of the DFA was further applied for the characterization of protein–protein interactions, especially antibody–antigen interactions . In particular, using a dsDNA molecule loaded in the unzip geometry, this MFS was used to implement a force‐based sandwich immunoassay in a double‐chip format (Figure C‐3) .…”

Section: First‐generation Molecular Force Sensorsmentioning

confidence: 99%

Molecular Force Sensors: From Fundamental Concepts toward Applications in Cell Biology

Göktas

Blank

2016

Adv Materials Inter

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Mechanical signals are central for the regulation of developmental, physiological, and pathological processes within biological systems. Force transduction across the cell–extracellular matrix (ECM) interface is highly crucial for regulating cell fate via mechanosensing and mechanotransduction cascades. The key molecules involved in these highly sophisticated processes have been identified in recent years. But little is still known about their interactions and in particular the molecular forces that determine these interactions. This is due to the limited availability of techniques that allow for investigating force propagation and mechanobiochemical signal conversion at the molecular level in live cells. In this progress report, currently available tools for measuring the molecular forces involved in cellular mechanosensing and mechanotransduction are summarized, specifically highlighting recent advances in the development of molecular force sensors (MFSs). MFSs convert the applied force into a fluorescence signal, allowing for a direct readout of tension with optical microscopy techniques. Moving from molecular design principles to applications of MFSs, important results are summarized, highlighting the new mechanistic information that has been obtained about mechanobiochemical processes at the cell–ECM interface. This progress report finishes with a critical discussion of current promises and limitations, providing perspectives for future research in this quickly evolving field

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Double chip protein arrays using recombinant single‐chain Fv antibody fragments

Cited by 15 publications

References 13 publications

Antibody Microarrays: Current Status and Key Technological Advances

Antibody Microarrays: Current Status and Key Technological Advances

Quantitative comparison of lipoprotein fractions derived from human plasma and serum by liquid chromatography-tandem mass spectrometry

Molecular Force Sensors: From Fundamental Concepts toward Applications in Cell Biology

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