Skip to main content

Protein Microarrays for Parasite Antigen Discovery

  • Protocol
  • First Online:
Parasite Genomics Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1201))

Abstract

The host serological profile to a parasitic infection, such as schistosomiasis, can be used to define potential vaccine and diagnostic targets. Determining the host antibody response using traditional approaches is hindered by the large number of putative antigens in any parasite proteome. Parasite protein microarrays offer the potential for a high-throughput host antibody screen to simplify this task. In order to construct the array, parasite proteins are selected from available genomic sequence and protein databases using bioinformatic tools. Selected open reading frames are PCR amplified, incorporated into a vector for cell-free protein expression, and printed robotically onto glass slides. The protein microarrays can be probed with antisera from infected/immune animals or humans and the antibody reactivity measured with fluorophore labeled antibodies on a confocal laser microarray scanner to identify potential targets for diagnosis or therapeutic or prophylactic intervention.

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 139.00
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. Vigil A, Davies DH, Felgner PL (2010) Defining the humoral immune response to infectious agents using high-density protein microarrays. Future Microbiol 5:241–251

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  2. Driguez P, Doolan DL, Loukas A, Felgner PL, McManus DP (2010) Schistosomiasis vaccine discovery using immunomics. Parasit Vectors 3:4

    Article  PubMed Central  PubMed  Google Scholar 

  3. Davies DH, Liang X, Hernandez JE, Randall A, Hirst S, Mu Y, Romero KM, Nguyen TT, Kalantari-Dehaghi M, Crotty S, Baldi P, Villarreal LP, Felgner PL (2005) Profiling the humoral immune response to infection by using proteome microarrays: high-throughput vaccine and diagnostic antigen discovery. Proc Natl Acad Sci U S A 102:547–552

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  4. Davies DH, Wyatt LS, Newman FK, Earl PL, Chun S, Hernandez JE, Molina DM, Hirst S, Moss B, Frey SE, Felgner PL (2008) Antibody profiling by proteome microarray reveals the immunogenicity of the attenuated smallpox vaccine modified vaccinia virus ankara is comparable to that of Dryvax. J Virol 82:652–663

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  5. Liu F, Lu J, Hu W, Wang SY, Cui SJ, Chi M, Yan Q, Wang XR, Song HD, Xu XN, Wang JJ, Zhang XL, Zhang X, Wang ZQ, Xue CL, Brindley PJ, McManus DP, Yang PY, Feng Z, Chen Z, Han ZG (2006) New perspectives on host-parasite interplay by comparative transcriptomic and proteomic analyses of Schistosoma japonicum. PLoS Pathog 2:e29

    Article  PubMed Central  PubMed  Google Scholar 

  6. Liu F, Zhou Y, Wang ZQ, Lu G, Zheng H, Brindley PJ, McManus DP, Blair D, Zhang QH, Zhong Y, Wang S, Han ZG, Chen Z (2009) The Schistosoma japonicum genome reveals features of host-parasite interplay. Nature 460:345–351

    Article  PubMed Central  Google Scholar 

  7. Chandra H, Reddy PJ, Srivastava S (2011) Protein microarrays and novel detection platforms. Expert Rev Proteomics 8:61–79

    Article  CAS  PubMed  Google Scholar 

  8. Austin J, Holway AH (2011) Contact printing of protein microarrays. Methods Mol Biol 785:379–394

    Article  CAS  PubMed  Google Scholar 

  9. He Y, Rappuoli R, De Groot AS, Chen RT (2010) Emerging vaccine informatics. J Biomed Biotechnol 2010:218590

    PubMed Central  PubMed  Google Scholar 

  10. Hoffmann KF, Fitzpatrick JM (2004) Gene expression studies using self-fabricated parasite cDNA microarrays. Methods Mol Biol 270:219–236

    CAS  PubMed  Google Scholar 

  11. Trieu A, Kayala MA, Burk C, Molina DM, Freilich DA, Richie TL, Baldi P, Felgner PL, Doolan DL (2011) Sterile protective immunity to malaria is associated with a panel of novel P. falciparum antigens. Mol Cell Proteomics 10(M111):007948

    PubMed  Google Scholar 

  12. Li MZ, Elledge SJ (2007) Harnessing homologous recombination in vitro to generate recombinant DNA via SLIC. Nat Methods 4:251–256

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We want to thank all the staff at Antigen Discovery Incorporated and the Protein Microarray Laboratory, University of California, Irvine. We also wish to thank Hamish McWilliam for his illustrations of schistosome lifecycle stages. PLF’s research was supported by National Institute of Allergy and Infectious Disease Grants U54AI065359 and U01AI078213. DLD, AL, and DPM receive support from the National Health and Medical Research Council of Australia.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Donald P. McManus .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2015 Springer Science+Business Media New York

About this protocol

Cite this protocol

Driguez, P. et al. (2015). Protein Microarrays for Parasite Antigen Discovery. In: Peacock, C. (eds) Parasite Genomics Protocols. Methods in Molecular Biology, vol 1201. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1438-8_13

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-1438-8_13

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-1437-1

  • Online ISBN: 978-1-4939-1438-8

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics