Comparison of Hydroxylated Print Additives on Antibody Microarray Performance

Wu, Peng; Grainger, David W.

doi:10.1021/pr060217d

Cited by 40 publications

(54 citation statements)

References 51 publications

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“…In fact, addition of other commonly used additives in printing buffer, glycerol or PEG, seemed to reduce attachment/immobilization. The results are consistent with Wu et al (Wu and Grainger, 2006) that among the various hydroxylated additives they investigated, PVA produced the most regular spot morphologies. The printing buffer also helps stabilize the droplet and ensures sufficient time for the attachment chemistry between peptides and surface to occur.…”

Section: Resultssupporting

confidence: 92%

Optimization of peptide arrays for studying antibodies to hepatitis C virus continuous epitopes

Ruwona

McBride

Chappel

et al. 2014

Journal of Immunological Methods

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Accurate and in-depth mapping of antibody responses is of great value in vaccine and antibody research. Using hepatitis C virus (HCV) as a model, we developed an affordable and high-throughput microarray-based assay for mapping antibody specificities to continuous antibody epitopes of HCV at high resolution. Important parameters in the chemistry for conjugating peptides/antigens to the array surface, the array layout, fluorophore choice and the methods for data analysis were investigated. Microscopic glass slide pre-coated with N-Hydroxysuccinimide (NHS)-ester (Slide H) was the preferred surface for conjugation of aminooxy-tagged peptides. This combination provides a simple chemical means to orient the peptides to the conjugation surface via an orthogonal covalent linkage at the N- or C-terminus of each peptide. The addition of polyvinyl alcohol to printing buffer gave uniform spot morphology, improved sensitivity and specificity of binding signals. Libraries of overlapping peptides covering the HCV E1 and E2 glycoprotein polypeptides (15-mer, 10 amino acids overlap) of 6 major HCV genotypes and the entire polypeptide sequence of the prototypic strain H77 were synthesized and printed in quadruplets in the assays. The utility of the peptide arrays were confirmed using HCV monoclonal antibodies (mAbs) specific to known continuous epitopes and immune sera of rabbits immunized with HCV antigens. The methods developed here can be easily adapted to studying antibody responses to antigens relevant in vaccine and autoimmune research.

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Section: Resultssupporting

confidence: 92%

Optimization of peptide arrays for studying antibodies to hepatitis C virus continuous epitopes

Ruwona

McBride

Chappel

et al. 2014

Journal of Immunological Methods

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“…For example, Wu and Grainger used different additives as a means to stabilize the cytokine antibodies for human IL-1β, IL-4, and TNF-α spotted onto a microarray [66]. With their approach, the chips were stable for 1 month at 4°C.…”

Section: Standard Cytokine Assaysmentioning

confidence: 99%

Bioanalytical chemistry of cytokines – A review

Stenken

Poschenrieder

2015

Analytica Chimica Acta

233

172

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Cytokines are bioactive proteins produced by many different cells of the immune system. Due to their role in different inflammatory disease states and maintaining homeostasis, there is enormous clinical interest in the quantitation of cytokines. The typical standard methods for quantitation of cytokines are immunoassay-based techniques including enzyme-linked immusorbent assays (ELISA) and bead-based immunoassays read by either standard or modified flow cytometers. A review of recent developments in analytical methods for measurements of cytokine proteins is provided. This review briefly covers cytokine biology and the analysis challenges associated with measurement of these biomarker proteins for understanding both health and disease. New techniques applied to immunoassay-based assays are presented along with the uses of aptamers, electrochemistry, mass spectrometry, optical resonator-based methods. Methods used for elucidating the release of cytokines from single cells as well as in vivo collection methods are described.

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“…[ 218 ] Additives in spotting solution can increase signal and improve storage. [ 219 ] These techniques are routinely used for low-cost mass-fabrication of immunoassay strip tests. A variety of chemicals can be patterned in multiple steps to produce complex architectures.…”

Section: Labeled Detection Of Analytesmentioning

confidence: 99%

Microfluidic Chips for Point‐of‐Care Immunodiagnostics

2011

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We might be at the turning point where research in microfluidics undertaken in academia and industrial research laboratories, and substantially sponsored by public grants, may provide a range of portable and networked diagnostic devices. In this Progress Report, an overview on microfluidic devices that may become the next generation of point-of-care (POC) diagnostics is provided. First, we describe gaps and opportunities in medical diagnostics and how microfluidics can address these gaps using the example of immunodiagnostics. Next, we conceptualize how different technologies are converging into working microfluidic POC diagnostics devices. Technologies are explained from the perspective of sample interaction with components of a device. Specifically, we detail materials, surface treatment, sample processing, microfluidic elements (such as valves, pumps, and mixers), receptors, and analytes in the light of various biosensing concepts. Finally, we discuss the integration of components into accurate and reliable devices.

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Comparison of Hydroxylated Print Additives on Antibody Microarray Performance

Cited by 40 publications

References 51 publications

Optimization of peptide arrays for studying antibodies to hepatitis C virus continuous epitopes

Optimization of peptide arrays for studying antibodies to hepatitis C virus continuous epitopes

Bioanalytical chemistry of cytokines – A review

Microfluidic Chips for Point‐of‐Care Immunodiagnostics

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