Simultaneous Detection of Multiple Cytokines from Conditioned Media and Patient's Sera by an Antibody-Based Protein Array System

Huang, Ruo‐Pan; Huang, Ruochun; Yan, Fan; Lin, Ying

doi:10.1006/abio.2001.5156

Cited by 231 publications

(121 citation statements)

References 26 publications

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“…Hueber et al [5] designed a synovial proteome microarray containing 650 candidate autoantigens, which reliably stratified patients with early rheumatoid arthritis into clinically relevant disease subsets. Huang et al [6] developed a cytokine array with increased sensitivity and a broader detection range compared with standard ELISAs. Hsueh et al [7] also compared the performance characteristics of autoantigen microarrays with those of commercial ELISAs in detecting five autoantibodies (anti-double-stranded DNA (dsDNA), Sm, RNP, Ro, La) in the sera of 80 SLE patients.…”

Section: Introductionmentioning

confidence: 99%

Customising an antibody leukocyte capture microarray for systemic lupus erythematosus: Beyond biomarker discovery

Lin

Adelstein

et al. 2010

Proteomics Clinical Apps

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Systemic lupus erythematosus (SLE) is a complex autoimmune disease that has heterogeneous clinical manifestation with diverse patterns of organ involvement, autoantibody profiles and varying degrees of severity of disease. Research and clinical experience indicate that different subtypes of SLE patients will likely benefit from more tailored treatment regimes, but we currently lack a fast and objective test with high enough sensitivity to enable us to perform such sub‐grouping for clinical use. In this article, we review how proteomic technologies could be used as such an objective test. In particular, we extensively review many leukocyte surface markers that are known to have an association with the pathogenesis of SLE, and we discuss how these markers can be used in the further development of a novel SLE‐specific antibody leukocyte capture microarray. In addition, we review some bioinformatics challenges and current methods for using the data generated by these cell‐capture microarrays in clinical use. In a broader context, we hope our experience in developing a disease specific cell‐capture microarray for clinical application can be a guide to other proteomic practitioners who intend to extend their technologies to develop clinical diagnostic and prognostic tests for complex diseases.

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Section: Introductionmentioning

confidence: 99%

Customising an antibody leukocyte capture microarray for systemic lupus erythematosus: Beyond biomarker discovery

Lin

Adelstein

et al. 2010

Proteomics Clinical Apps

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“…Various detection methods have been used: rolling circle amplification (10,11), resonance light scattering (12), enhanced chemiluminescence (13), tyramide signal amplification (14), and fluorescence (15,16). Huang et al (13) used arrayed sandwich assays to detect 24 different cytokines in conditioned media and serum samples. The antibodies were spotted onto membranes, and after incubation of samples on the membranes, bound antigens were detected by a mixture of biotinylated detection antibodies.…”

Section: Studies On Bodily Fluids or Secreted Proteinsmentioning

confidence: 99%

“…Tumor intersti-tial fluid samples were collected from breast tumor tissue and normal surrounding breast tissue and analyzed by mass spectrometry, Western immunoblotting, and cytokine-specific antibody microarrays (19). The microarrays were similar to those developed by Huang et al (13), described above, and targeted 70 different cytokines. The characterization of cytokine levels in the fluid will be used with the other types of proteomic data to elucidate the interactions between the cells in the tumor environment.…”

Section: Studies On Bodily Fluids or Secreted Proteinsmentioning

confidence: 99%

“…The cytokine arrays developed by Huang et al (13), as described above, were applied to the measurement of 35 different cytokines in lysates from cultured breast cancer cells and endometriosis tissue (23). Another study used two-color comparative fluorescence to examine the effect of UV irradiation on protein expression in apoptosis signaling pathways (24).…”

Section: Tissue and Cell Culture Studiesmentioning

confidence: 99%

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Antibody Arrays in Cancer Research

Haab

2005

Molecular & Cellular Proteomics

222

131

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Antibody microarrays enable the parallel detection of multiple proteins in low sample volumes. Continued improvements in the technology have increased the usefulness of the methods for basic and applied biological research, including cancer research. Several different versions of antibody microarray formats and methods have been developed, each with various advantages, disadvantages, and optimal applications. This review looks at the applications of those various technologies to cancer research.The experimental features of microarrays have advantages for cancer research. The low sample volumes result in the consumption of small amounts of both precious clinical samples and expensive antibodies. The assays can be run efficiently in parallel, enabling studies on the large populations of samples that are necessary for marker discovery and validation. In addition, the assays can have good reproducibility, high sensitivity, and quantitative accuracy over large concentration ranges (1). Antibody and protein arrays are complementary and in some aspects preferable to separation-based and mass spectrometry-based technologies. Reproducibility and throughput can be higher, and the identities of the measured proteins are known or can be readily characterized.Therefore, specific hypotheses regarding the nature of molecular alterations can be tested and generated, and the observed measurements can be biologically interpreted.A common application of antibody arrays in cancer research is the identification of biomarkers or molecules that are potentially valuable for diagnosis or prognosis or as surrogate markers of drug response. The multiplex capability of antibody arrays allows the efficient screening of many marker candidates to reveal associations between proteins and disease states or experimental conditions. Multiplexed measurements also allow the evaluation of the use of multiple markers in combination. The use of combinations of proteins for disease diagnostics may produce fewer false positive and false negative results as compared with tests based on single proteins. Antibody microarrays, by increasing the number of proteins that can be conveniently measured in clinical samples, could more significantly take advantage of the benefit of using combined markers in diagnostics. Other example applications of antibody microarrays in cancer research are to evaluate the coordinated changes of members of signaling pathways or to measure changes in expression levels of a class of proteins, such as angiogenesis factors.This review covers the major applications of antibody array technologies to cancer research. The first section gives an overview of the main features of the various antibody microarray technologies. Details of the technologies will not be covered extensively; reviews with a greater technological focus are available elsewhere (2-6). The next two sections cover applications to cancer research and are divided according to the type of sample analyzed: fluids (second section) and cells (third section). The final section...

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“…Schweitzer and colleagues [9,10] described a new methodology named Rolling Circle Amplification for signal amplification on microarrays, which relies on enzymatic extension of an primer-antibody conjugate followed by hybridization of labeled probes to the generated DNA strand. As an alternative approach to fluorescent detection, Huang and colleagues [29] applied chemiluminescence for the sensitive detection of multiple cytokines. However, all currently presented solutions either lack the possibility of multiplex screening of analytes against enzymes, or require complicated liquid handling, surface modifications, and additional equipment.…”

Section: Introductionmentioning

confidence: 99%

Subnanoliter enzymatic assays on microarrays

et al. 2005

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Many areas of research today are based on enzymatic assays most of which are still performed as enzyme-linked immunosorbent assays in microtiter plates. The demand for highly parallel screening of thousands of samples eventually led to a miniaturization and automation of these assays. However, the final transfer of enzymatic assays from a microtiter-based technology to microarrays has proven to be difficult for various reasons, such as the inability to maintain unbound reaction products on the spot of reaction or the missing capability of multiplexing. Here, we have conducted multiplex enzymatic assays in subnanoliter volumes on a single microarray using the multiple spotting technology. We were able to measure enzymatic activity with a sensitivity down to 35 enzyme molecules, applying only conventional flat microarray surfaces and standard microarray hardware. We have performed assays of inhibition and applied this format for the detection of prognostic markers, such as cathepsin D. The new approach allows the rapid and multiplex screening of thousands of samples on a single microarray with applications in drug screening, metagenomics, and high-throughput enzyme assays.

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Simultaneous Detection of Multiple Cytokines from Conditioned Media and Patient's Sera by an Antibody-Based Protein Array System

Cited by 231 publications

References 26 publications

Customising an antibody leukocyte capture microarray for systemic lupus erythematosus: Beyond biomarker discovery

Customising an antibody leukocyte capture microarray for systemic lupus erythematosus: Beyond biomarker discovery

Antibody Arrays in Cancer Research

Subnanoliter enzymatic assays on microarrays

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