Antibody Detection in Human Serum Using a Versatile Protein Chip Platform Constructed by Applying Nanoscale Self-Assembled Architectures on Gold

Pavlickova, Petra; Jensen, Nina Mejlhede; Paul, Heiko; Schaeferling, Michael; Giammasi, Chiara; Kruschina, Margit; Du, Wei‐Dong; Theisen, Michael; Ibba, Michael; Ortigao, Flavio; Kambhampati, Dev

doi:10.1021/pr0200036

Cited by 46 publications

(21 citation statements)

References 23 publications

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“…Regulating biomolecular immobilization is essential for chemical modification of biochips and for biosensing of surfaces for protein capture . We used thin film gold surface implanted on glass, which had 192 spots available as the parent substrate due to their stability and chemical assembly features through Au‐S bonds in biochips (supplied by Thermohybaid, Interactiva Division, Ulm, Germany) …”

Section: Methodsmentioning

confidence: 99%

S‐S‐PEG‐COOH Self‐Assembled Monolayer on Gold Surface Enabled a Combined Assay for Serological EBV Antibody Isotypes

Liu

et al. 2018

Proteomics Clinical Apps

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Purpose: Epstein-Barr virus (EBV) is a ubiquitous human gamma herpes virus that infects human epithelial cells and B lymphocytes. It would be potentially valuable to develop novel combined assays to benefit screening for large panels of samples of EBV infectious diseases. Experimental design: A simple antigen-probed biochip that is modified with S-S-PEG-COOH and is used as a label-free high-throughput screening method for a combined detection of EBV capsid antigen IgM antibody, capsid antigen IgG antibody, and nuclear antigen IgG antibody.Results: This protein biochip has similar feasibility, sensitivity, and specificity in comparison with Liaison chemiluminescent immunoassay (CLIA). Detection limit of the EBV antibodies by the biochip is almost identical to that by CLIA-L (2.91 U mL -1 vs 3.00 U mL -1 for EBNA-1 IgG, 8 U mL -1 vs10 U mL -1 for EBV-VCA IgG, and 3.5 U mL -1 vs 10 U mL -1 for EBV-VCA IgM). Tests of the three serological antibodies against EBV by the biochip are consistent with the CLIA-L method in 274 clinical sera, respectively. Finally, the combined biochip is successfully utilized for diagnostic identification of EBV infection in 14 patients with infectious mononucleosis (IM) and 25 patients with systemic lupus erythematosus SLE, as well as additional 10 known real-time PCR positive patients.

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Section: Methodsmentioning

confidence: 99%

S‐S‐PEG‐COOH Self‐Assembled Monolayer on Gold Surface Enabled a Combined Assay for Serological EBV Antibody Isotypes

Liu

et al. 2018

Proteomics Clinical Apps

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“…Other array formats used for serum protein analysis include selfassembled monolayer array, filtration-based array, multiplexed photoaptamer-based arrays, phage-based array, and RP array [135][136][137][138][139]. The filtration-based protein array can improve the overall reaction kinetic rate by ten-fold and enhance the sensitivity and specificity of the assay.…”

Section: Protein Arraysmentioning

confidence: 99%

Human body fluid proteome analysis

2006

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The focus of this article is to review the recent advances in proteome analysis of human body fluids, including plasma/serum, urine, cerebrospinal fluid, saliva, bronchoalveolar lavage fluid, synovial fluid, nipple aspirate fluid, tear fluid, and amniotic fluid, as well as its applications to human disease biomarker discovery. We aim to summarize the proteomics technologies currently used for global identification and quantification of body fluid proteins, and elaborate the putative biomarkers discovered for a variety of human diseases through human body fluid proteome (HBFP) analysis. Some critical concerns and perspectives in this emerging field are also discussed. With the advances made in proteomics technologies, the impact of HBFP analysis in the search for clinically relevant disease biomarkers would be realized in the future.

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“…We were able to detect femtomoles of target, which is comparable to the dot-ELISA [46]. A lower analytical sensitivity may be feasible by optimizing the antigen-biochip linkage method to maintain antigen conformation and optimal antibody-antigen interactions [47]. Further application of the GC-FP biochip for multiplexed serum antibody detection involves measuring how each antigen interacts with individual immune responses, which heterogeneously produce antibodies that bind various epitopes on B. burgdorferi proteins during infection [48,49].…”

Section: Quantitative Analysis Using the Gc-fp Biochip Is Highly Sensmentioning

confidence: 92%

A fluorescent plasmonic biochip assay for multiplex screening of diagnostic serum antibody targets in human Lyme disease

et al. 2020

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Lyme disease (LD) diagnosis using the current two-tier algorithm is constrained by low sensitivity for early-stage infection and ambiguity in determining treatment response. We recently developed a protein microarray biochip that measures diagnostic serum antibody targets using grating-coupled fluorescent plasmonics (GC-FP) technology. This strategy requires microliters of blood serum to enable multiplexed biomarker screening on a compact surface and generates quantitative results that can be further processed for diagnostic scoring. The GC-FP biochip was used to detect serum antibodies in patients with active and convalescent LD, as well as various negative controls. We hypothesized that the quantitative, high-sensitivity attributes of the GC-FP approach permit: 1) screening of antibody targets predictive for LD status, and 2) development a diagnostic algorithm that is more sensitive, specific, and informative than the standard ELISA and Western blot assays. Notably, our findings led to a diagnostic algorithm that may be more sensitive than the current standard for detecting early LD, while maintaining 100% specificity. We further show that analysis of relative antibody levels to predict disease status, such as in acute and convalescent stages of infection, is possible with a highly sensitive and quantitative platform like GC-FP. The results from this study add to the urgent conversation regarding better diagnostic strategies and more effective treatment for patients affected by tick-borne disease.

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Antibody Detection in Human Serum Using a Versatile Protein Chip Platform Constructed by Applying Nanoscale Self-Assembled Architectures on Gold

Cited by 46 publications

References 23 publications

S‐S‐PEG‐COOH Self‐Assembled Monolayer on Gold Surface Enabled a Combined Assay for Serological EBV Antibody Isotypes

S‐S‐PEG‐COOH Self‐Assembled Monolayer on Gold Surface Enabled a Combined Assay for Serological EBV Antibody Isotypes

Human body fluid proteome analysis

A fluorescent plasmonic biochip assay for multiplex screening of diagnostic serum antibody targets in human Lyme disease

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