Advertisement

Neuroproteomic Profiling of Cerebrospinal Fluid (CSF) by Multiplexed Affinity Arrays

  • Anna Häggmark-Månberg
  • Peter Nilsson
  • Jochen M. Schwenk
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1598)

Abstract

Protein profiling through affinity proteomic approaches represents a powerful strategy for the analysis of human body fluids. Cerebrospinal fluid (CSF), being the fluid proximal to the central nervous system, is commonly analyzed in the context of neurological diseases, and can offer novel insights into the physiological state of the brain. Ultimately, and by analyzing the presence of brain-derived proteins in larger sets of samples that represent different phenotypes, profiling of CSF may serve as an important source to discover and verify disease-associated markers. Here, we describe a multiplexed and flexible protein profiling approach using antibody-based assays on suspension bead arrays. Through streamlined sample processing, protein biotinylation, and single-binder assay readout, this method enables high-throughput neuroproteomic analysis of up to 384 proteins in 384 samples.

Key words

Suspension bead array Antibody Microarray Cerebrospinal fluid Direct labeling Protein profiling 

Notes

Acknowledgments

We like to thank the entire staff of the Human Protein Atlas (HPA) for their efforts. This work was funded by the PRONOVA project (VINNOVA, Swedish Governmental Agency for Innovation Systems), and by grants from the Knut and Alice Wallenberg Foundation and Science for Life Laboratory, Stockholm.

References

  1. 1.
    Hanash S (2003) Disease proteomics. Nature 422:226–232CrossRefPubMedGoogle Scholar
  2. 2.
    Ayoglu B, Chaouch A, Lochmuller H, Politano L, Bertini E, Spitali P, Hiller M, Niks EH, Gualandi F, Ponten F, Bushby K, Aartsma-Rus A, Schwartz E, Le Priol Y, Straub V, Uhlen M, Cirak S, t Hoen PA, Muntoni F, Ferlini A, Schwenk JM, Nilsson P, Al-Khalili Szigyarto C (2014) Affinity proteomics within rare diseases: a BIO-NMD study for blood biomarkers of muscular dystrophies. EMBO Mol Med 6:918–936CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Häggmark A, Mikus M, Mohsenchian A, Hong M-G, Forsström B, Gajewska B, Barańczyk-Kuźma A, Uhlén M, Schwenk JM, Kuźma–Kozakiewicz M, Nilsson P (2014) Plasma profiling reveals three proteins related to amyotrophic lateral sclerosis. Ann Clin Transl Neurol 1(8):544–553CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Neiman M, Hedberg JJ, Donnes PR, Schuppe-Koistinen I, Hanschke S, Schindler R, Uhlen M, Schwenk JM, Nilsson P (2011) Plasma profiling reveals human fibulin-1 as candidate marker for renal impairment. J Proteome Res 10:4925–4934CrossRefPubMedGoogle Scholar
  5. 5.
    Schwenk JM, Gry M, Rimini R, Uhlen M, Nilsson P (2008) Antibody suspension bead arrays within serum proteomics. J Proteome Res 7:3168–3179CrossRefPubMedGoogle Scholar
  6. 6.
    Hokfelt T, Broberger C, Xu ZQ, Sergeyev V, Ubink R, Diez M (2000) Neuropeptides—an overview. Neuropharmacology 39:1337–1356CrossRefPubMedGoogle Scholar
  7. 7.
    Haggmark A, Bystrom S, Ayoglu B, Qundos U, Uhlen M, Khademi M, Olsson T, Schwenk JM, Nilsson P (2013) Antibody-based profiling of cerebrospinal fluid within multiple sclerosis. Proteomics 13:2256–2267CrossRefPubMedGoogle Scholar
  8. 8.
    Schwenk JM, Igel U, Neiman M, Langen H, Becker C, Bjartell A, Ponten F, Wiklund F, Gronberg H, Nilsson P, Uhlen M (2010) Toward next generation plasma profiling via heat-induced epitope retrieval and array-based assays. Mol Cell Proteomics 9:2497–2507CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  • Anna Häggmark-Månberg
    • 1
  • Peter Nilsson
    • 1
  • Jochen M. Schwenk
    • 1
  1. 1.Affinity Proteomics, Science for Life Laboratory, School of BiotechnologyKTH - Royal Institute of TechnologySolnaSweden

Personalised recommendations