Skip to main content

Detecting and Quantifying Multiple Proteins in Clinical Samples in High-Throughput Using Antibody Microarrays

  • Protocol
  • First Online:
Protein Microarray for Disease Analysis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 723))

Abstract

Many diagnostic and prognostic tests performed in the clinic today rely on the sensitive detection and quantification of a single protein, usually by means of an immunoassay. Even in the case of monogenic diseases, however, single markers are often insufficient to provide highly reliable predictions of disease onset, and the accuracy of these predictions only decreases for polygenic diseases and for very early detection or prediction. Recent studies have shown that predictive reliability increases dramatically when multiple markers are analyzed simultaneously. Antibody microarrays provide a powerful way to quantify the abundance of many different proteins simultaneously in a variety of sample types, including serum, urine, and tissue explants. Because the assay is highly miniaturized, very little sample is required and the assay can be performed in high-throughput. Using antibody microarrays, we have been able to identify prognostic markers of early mortality in patients with end-stage renal disease and have built multivariate models based on these markers. We anticipate that antibody microarrays will prove similarly useful in other discovery-based efforts and may ultimately enjoy routine use in clinical labs.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 159.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Alizadeh AA, Eisen MB, Davis RE et al (2000) Distinct types of diffuse large B-cell lymphoma identified by gene expression profiling. Nature 403:503–511

    Article  PubMed  CAS  Google Scholar 

  2. Liang Y, Diehn M, Watson N et al (2005) Gene expression profiling reveals molecularly and clinically distinct subtypes of glioblastoma multiforme. Proc Natl Acad Sci U S A 102:5814–5819

    Article  PubMed  CAS  Google Scholar 

  3. Ramaswamy S, Tamayo P, Rifkin R et al (2001) Multiclass cancer diagnosis using tumor gene expression signatures. Proc Natl Acad Sci U S A 98:15149–15154

    Article  PubMed  CAS  Google Scholar 

  4. Zhou SF, Di YM, Chan E et al (2008) Clinical pharmacogenetics and potential application in personalized medicine. Curr Drug Metab 9:738–784

    Article  PubMed  CAS  Google Scholar 

  5. Duffy MJ, Crown J (2008) A personalized approach to cancer treatment: how biomarkers can help. Clin Chem 54:1770–1779

    Article  PubMed  CAS  Google Scholar 

  6. Hanash S (2003) Disease proteomics. Nature 422:226–232

    Article  PubMed  CAS  Google Scholar 

  7. Schweitzer B, Roberts S, Grimwade B et al (2002) Multiplexed protein profiling on microarrays by rolling-circle amplification. Nat Biotechnol 20:359–365

    Article  PubMed  CAS  Google Scholar 

  8. Shao W, Zhou Z, Laroche I et al (2003) Optimization of rolling-circle amplified protein microarrays for multiplexed protein profiling. J Biomed Biotechnol 5:299–307

    Article  Google Scholar 

  9. Knickerbocker T, Chen JR, Thadhani R, MacBeath G (2007) An integrated approach to prognosis using protein microarrays and nonparametric methods. Mol Syst Biol 3(123):1–8

    Google Scholar 

  10. MacBeath G (2002) Protein microarrays and proteomics. Nat Genet 32:526–532

    Article  PubMed  CAS  Google Scholar 

  11. Li C, Simeone DM, Brenner DE et al (2009) Pancreatic cancer serum detection using a lectin/glyco-antibody array method. J Proteome Res 8:483–492

    Article  PubMed  CAS  Google Scholar 

  12. Belov L, Mulligan SP, Barber N et al (2006) Analysis of human leukaemias and lymphomas using extensive immunophenotypes from an antibody microarray. Br J Haematol 135:184–197

    Article  PubMed  CAS  Google Scholar 

  13. Thadhani R, Tonelli M (2006) Cohort studies: marching forward. Clin J Am Soc Nephrol 1:1117–1123

    Article  PubMed  Google Scholar 

  14. USRSD (2005) National Institutes of Health. National Institute of Diabetes and Digestive and Kidney Diseases, Bethesda

    Google Scholar 

  15. Buja A, Hastie T, Tibshirani R (1989) Linear smoothers and additive models (with discussion). Ann Statist 17:453–555

    Article  Google Scholar 

  16. Natarajan M, Lin KM, Hsueh RC et al (2006) A global analysis of cross-talk in a mammalian cellular signalling network. Nat Cell Biol 8:571–580

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgment

We thank Ravi Thadhani for directing ArMORR (Accelerated Mortality on Renal Replacement), a prospective study of ESRD patients, and Jiunn-Ren Chen for data analysis and interpretation. This work was supported by awards from the WM Keck Foundation and the Arnold and Mabel Beckman Foundation, and by grants from the National Institutes of Health (DK071674 and DK068465). T.K. is the recipient of an Eli Lilly Graduate Student Fellowship.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Tanya Knickerbocker .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2011 Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Knickerbocker, T., MacBeath, G. (2011). Detecting and Quantifying Multiple Proteins in Clinical Samples in High-Throughput Using Antibody Microarrays. In: Wu, C. (eds) Protein Microarray for Disease Analysis. Methods in Molecular Biology, vol 723. Humana Press. https://doi.org/10.1007/978-1-61779-043-0_1

Download citation

  • DOI: https://doi.org/10.1007/978-1-61779-043-0_1

  • Published:

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-61779-042-3

  • Online ISBN: 978-1-61779-043-0

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics