Jump to ContentJump to Main Navigation
Show Summary Details
More options …

Clinical Chemistry and Laboratory Medicine (CCLM)

Published in Association with the European Federation of Clinical Chemistry and Laboratory Medicine (EFLM)

Editor-in-Chief: Plebani, Mario

Ed. by Gillery, Philippe / Greaves, Ronda / Lackner, Karl J. / Lippi, Giuseppe / Melichar, Bohuslav / Payne, Deborah A. / Schlattmann, Peter

IMPACT FACTOR 2018: 3.638

CiteScore 2018: 2.44

SCImago Journal Rank (SJR) 2018: 1.191
Source Normalized Impact per Paper (SNIP) 2018: 1.205

See all formats and pricing
More options …
Volume 41, Issue 2


Detection of Multiple Cytokines by Protein Arrays from Cell Lysate and Tissue Lysate

Ying Lin / Ruochun Huang / Xuan Cao / Shen-Ming Wang / Qing Shi / Ruo-Pan Huang
Published Online: 2005-06-01 | DOI: https://doi.org/10.1515/CCLM.2003.023


Previously we demonstrated that multiple cytokines could be simultaneously detected using an antibody-based protein array system with high sensitivity and specificity from conditioned medium and serum. Here, we created a higher density array system to simultaneously detect 35 cytokines from cell lysates and tissue lysates. This assay combines the advantages of the specificity of enzyme-linked immunosorbent assays (ELISA), sensitivity of enhanced chemiluminescence (ECL), and high-throughput of microspot. In this system, capture antibodies dissolved in methanol were spotted onto polyvinylidene difluoride (PVDF) membranes. The membranes were then incubated with tissue lysates or cell lysates. After removing unbound proteins by extensive washing, the membranes were exposed to horseradish peroxidase (HRP)-conjugated antibody(ies). The signals were visualized with an ECL system. High specificity, sensitivity, and accuracy of this approach were demonstrated. This approach can be used in any general laboratory setting without any sophisticated equipment. It should be feasible to extend this concept to develop a high-throughput protein array system. Combining nitrocellulose membrane-based and PVDF membrane-based approaches, the human cytokine array system can be applied to detect multiple cytokine expression from cell lysate, tissue lysate, serum, plasma, and conditioned medium. Future applications of this new approach include direct protein expression profiling, immunological disease diagnostics, and discovery of new biomarkers.

About the article

Published Online: 2005-06-01

Published in Print: 2003-02-21

Citation Information: Clinical Chemistry and Laboratory Medicine, Volume 41, Issue 2, Pages 139–145, ISSN (Print) 1434-6621, DOI: https://doi.org/10.1515/CCLM.2003.023.

Export Citation

Citing Articles

Here you can find all Crossref-listed publications in which this article is cited. If you would like to receive automatic email messages as soon as this article is cited in other publications, simply activate the “Citation Alert” on the top of this page.

Wei Huang, Kelly Whittaker, Huihua Zhang, Jian Wu, Si-Wei Zhu, and Ruo-Pan Huang
ASSAY and Drug Development Technologies, 2018
Brian B. Haab
Molecular & Cellular Proteomics, 2005, Volume 4, Number 4, Page 377
Getulio Nogueira-Filho, Igor Pesun, Cindy Isaak-Ploegman, Manjula Wijegunasinghe, Tom Wierzbicki, and Christopher A. McCulloch
Journal of Periodontology, 2014, Volume 85, Number 11, Page 1582
T S Gujral, R L Karp, A Finski, M Chan, P E Schwartz, G MacBeath, and P Sorger
Oncogene, 2013, Volume 32, Number 29, Page 3470
Ching-Liang Ho, Terra L. Lasho, Joseph H. Butterfield, and Ayalew Tefferi
Leukemia Research, 2007, Volume 31, Number 10, Page 1389
I Leonard Bernstein, James T. Li, David I. Bernstein, Robert Hamilton, Sheldon L. Spector, Ricardo Tan, Scott Sicherer, David B.K. Golden, David A. Khan, Richard A. Nicklas, Jay M. Portnoy, Joann Blessing-Moore, Linda Cox, David M. Lang, John Oppenheimer, Christopher C. Randolph, Diane E. Schuller, Stephen A. Tilles, Dana V. Wallace, Estelle Levetin, and Richard Weber
Annals of Allergy, Asthma & Immunology, 2008, Volume 100, Number 3, Page S1
Se Hoon Choi, Karthikeyan Veeraraghavalu, Orly Lazarov, Seoan Marler, Richard M. Ransohoff, Jan Marino Ramirez, and Sangram S. Sisodia
Neuron, 2008, Volume 59, Number 4, Page 568
Chaya Moroz, Anna Grunspan, Muayad A. Zahalka, Leonid Traub, Yona Kodman, and Isaac Yaniv
Experimental Hematology, 2006, Volume 34, Number 2, Page 159
Paul J Utz
Clinical Immunology, 2004, Volume 111, Number 2, Page 153
J. Hu, X. He, K. A. Baggerly, K. R. Coombes, B. T.J. Hennessy, and G. B. Mills
Bioinformatics, 2007, Volume 23, Number 15, Page 1986
Stephen R. Master, Charlene Bierl, and Larry J. Kricka
Drug Discovery Today, 2006, Volume 11, Number 21-22, Page 1007
Mohamed Saiel Saeed Alhamdani, Christoph Schröder, and Jörg D. Hoheisel
PROTEOMICS, 2010, Volume 10, Number 17, Page 3203
Aldo Roda, Massimo Guardigli, Patrizia Pasini, Mara Mirasoli, Elisa Michelini, and Monica Musiani
Analytica Chimica Acta, 2005, Volume 541, Number 1-2, Page 25
Ruo-Pan Huang
Expert Review of Proteomics, 2007, Volume 4, Number 2, Page 299
James F. Rusling, Challa V. Kumar, J. Silvio Gutkind, and Vyomesh Patel
The Analyst, 2010, Volume 135, Number 10, Page 2496
Marta Sanchez-Carbayo
Expert Opinion on Medical Diagnostics, 2008, Volume 2, Number 3, Page 249
Larry J. Kricka and Stephen R. Master
Molecular Biotechnology, 2008, Volume 38, Number 1, Page 19
Hisashi Koga, Motoki Kyo, Kazue Usui-Aoki, and Kazuki Inamori
ELECTROPHORESIS, 2006, Volume 27, Number 18, Page 3676
Paul J. Utz
Immunological Reviews, 2005, Volume 204, Number 1, Page 264
Richard Barry and Mikhail Soloviev
PROTEOMICS, 2004, Volume 4, Number 12, Page 3717

Comments (0)

Please log in or register to comment.
Log in