Advertisement

Analytical and Bioanalytical Chemistry

, Volume 405, Issue 12, pp 3973–3982 | Cite as

Design and application of GB virus C (GBV-C) peptide microarrays for diagnosis of GBV-C/HIV-1 co-infection

  • Leticia Fernández
  • M. José Bleda
  • M. José Gómara
  • Isabel HaroEmail author
Original Paper

Abstract

The main objectives of the design of GB virus C (GBV-C) peptide microarrays are the miniaturisation of antigen–antibody interaction assays, the simultaneous analysis of several peptide sequences and the reduction in the volume of serum required from patients since this always represents a limiting factor in studies to develop new systems for diagnosing human diseases. We herein report the design of a microarray immunoassay based on synthetic peptides derived from the GBV-C E2 protein to evaluate their diagnostic value in detecting anti-E2 antibodies in HIV-1 patients. To this end, peptide microarrays were initially prepared to identify the most relevant epitopes in the GBV-C E2 protein. Thus, 124 peptides composed of 18 amino acids covering the whole E2-protein sequence, with 15 residue overlaps, were spotted in triplicate onto γ-aminopropyl silane-functionalised adsorbent binding slides. The procedure to select the E2 protein epitopes was carried out using serum samples from HIV-1-infected patients. The samples had previously been tested for the presence or absence of GBV-C anti-E2 antibodies by means of the Abbott test. Thus, 11 specific epitopes in the GBV-C E2 protein were identified. Subsequently, peptide antigen microarrays were constructed using the E2 epitopes identified to detect GBV-C anti-E2 antibodies in the serum of HIV-1-infected patients with no known GBV-C co-infection. The 11 peptides selected identified anti-E2 GBV-C antibodies among HIV-1-infected patients, and a reactivity of 47 % was established. The potential antigenic peptides selected could be considered a useful tool for designing a new diagnostic system based on peptide microarrays to determine anti-GBV-C E2 antibodies in the serum of HIV-1-infected patients.

Keywords

GBV-C/HIV-1 co-infection Diagnosis GBV-C peptides Microarrays 

Notes

Acknowledgments

The authors greatly acknowledge Professors Tillmann (Duke Clinical Research Institute, USA), Ercilla (Hospital Clinic, Barcelona, Spain) and Casanova (Hospital de Bellvitge, Barcelona, Spain) for the donation of the serum samples. We also acknowledge Drs. Lidia Sevilla and Ignacio Pons (Scientific Parc of Barcelona, Spain) for their helpful advice during the preparation and processing of microarrays. This work was funded by grant CTQ2009-13969-C02-01 from the Spanish Ministerio de Ciencia e Innovación. LF is a recipient of a FPI grant from the Spanish Ministerio de Ciencia e Innovación.

Supplementary material

216_2012_6585_MOESM1_ESM.pdf (331 kb)
ESM 1 (PDF 330 kb)

References

  1. 1.
    Baggio-Zappia GL, Granato CFH (2009) Clin Chem Lab Med 47:12–19CrossRefGoogle Scholar
  2. 2.
    George S, Varmaz D, Stapleton JT (2006) J Infect Dis 193:451–454Google Scholar
  3. 3.
    Mphahlele MJ, Lau GKK, Carman WF (1998) Liver 18:143–155Google Scholar
  4. 4.
    Theodore D, Lemon SM (1997) Hepatology 25:1285–1286CrossRefGoogle Scholar
  5. 5.
    Heringlake S, Ockenga J, Tillmann HL, Trautwein C, Meissner D, Stoll M, Hunt J, Jou C, Solomon N, Schmidt RE, Manns MP (1998) J Infect Dis 177:1723–1726CrossRefGoogle Scholar
  6. 6.
    Tillmann HL, Heiken H, Knapik-Botor A, Heringlake S, Ockenga J, Wilber JC, Goergen B, Detmer J, McMorrow M, Stoll M, Schmidt RE, Manns MP (2001) N Engl J Med 345:715–724CrossRefGoogle Scholar
  7. 7.
    Xiang JH, Wunschmann S, Diekema DJ, Klinzman D, Patrick KD, George SL, Stapleton JT (2001) N Engl J Med 345:707–714CrossRefGoogle Scholar
  8. 8.
    Zhang W, Chaloner K, Tillmann HL, Williams CF, Stapleton JT (2006) HIV Medicine 7:173–180CrossRefGoogle Scholar
  9. 9.
    Williams CF, Klinzman D, Yamashita TE, Xiang JH, Polgreen PM, Rinaldo C, Liu C, Phair J, Margolick JB, Zdunek D, Hess G, Stapleton JT (2004) N Engl J Med 350:981–990Google Scholar
  10. 10.
    Van der Bij AK, Kloosterboer N, Prins M, Boeser-Nunnink B, Geskus RB, Lange JMA, Coutinho RA, Schuitemaker H (2005) J Infect Dis 191:678–685CrossRefGoogle Scholar
  11. 11.
    Künkel U, Höhne M, Berg T, Hopf U, Kekulé AS, Frösner G, Pauli G, Schreier E (1998) J Hepatol 28:978–984CrossRefGoogle Scholar
  12. 12.
    Tacke M, Kiyosawa K, Stark K, Schlueter V, Ofenloch-Haehnle B, Hess G, Engel AM (1997) Lancet 349:318–320Google Scholar
  13. 13.
    Reshetnyak V, Karlovich T, Ilchenko L (2008) World J Gastroenterol 14:4725–4734CrossRefGoogle Scholar
  14. 14.
    Thomas DL, Vlahov D, Alter HJ, Hunt JC, Marshall R, Astemborski J, Nelson KE (1998) J Infect Dis 177:539–542CrossRefGoogle Scholar
  15. 15.
    Toniutto P, Fabris C, Barbone F, Tisminetzky SG, Liani D, Galai T, Barillari G, Biffoni F, Gasparini V, Pirisi M (1999) Clin Diagn Lab Immunol 6:573–576Google Scholar
  16. 16.
    Berzsenyi MD, Roberts SK (2006) J Infect Dis 194:407–409CrossRefGoogle Scholar
  17. 17.
    Herrera E, Tenckhoff S, Gomara M, Galatola R, Bleda M, Gil C, Ercilla G, Gatell JM, Tillmann HL, Haro I (2010) J Med Chem 53:6054–6063Google Scholar
  18. 18.
    Uttamchandani M, Yao SQ (2008) Curr Pharm Design 14:2428–2438Google Scholar
  19. 19.
    Foong YM, Fu JQ, Yao SQ, Uttamchandani M (2012) Curr Opinion Chem Biol 16:234–242Google Scholar
  20. 20.
    Andresen H, Grötzinger C, Zarse K, Kreuzer OJ, Ehrentreich-Förster E, Bier FF (2006) Proteomics 6:1376–1384CrossRefGoogle Scholar
  21. 21.
    Hueber W, Kidd BA, Tomooka BH, Lee BJ, Bruce B, Fries J, Sonderstrup G, Monach P, Drijfhout J, van Venrooij W, Utz P, Genovese M, Robinson W (2005) Arthritis Rheum 52:2645–2655Google Scholar
  22. 22.
    Hecker M, Lorenz P, Steinbeck F, Hong L, Riemekasten G, Li YX, Zettl UK, Thiesen HJ (2012) Autoimmun Rev 11:180–190Google Scholar
  23. 23.
    Wilcoxon F (1945) Biom Bull 1:80–83CrossRefGoogle Scholar
  24. 24.
    Kruskal WH (1957) J Am Stat Assoc 52:356–360CrossRefGoogle Scholar
  25. 25.
    Benjamini Y, Hochberg Y (1995) J R Stat Soc Series B (Methodological) 57:289–300Google Scholar
  26. 26.
    Simes RJ (1986) Biometrika 73:751–754CrossRefGoogle Scholar
  27. 27.
    Benjamini Y, Yekutieli D (2001) Ann Stat 29:1165–1188CrossRefGoogle Scholar
  28. 28.
    Delong ER, Delong DM, Clarkepearson DI (1988) Biometrics 44:837–845CrossRefGoogle Scholar
  29. 29.
    Hanley JA, McNeil BJ (1982) Radiology 143:29–36Google Scholar
  30. 30.
    Hanley JA, McNeil BJ (1983) Radiology 148:839–843Google Scholar
  31. 31.
    StataCorp (2011) Stata Statistical Software: Release 12. StataCorp LP, College Station, TXGoogle Scholar
  32. 32.
    Cretich M, Damin F, Pirri G, Chiari M (2006) Biomol Eng 23:77–88Google Scholar
  33. 33.
    Gómara MJ, Fernández L, Pérez T, Tenckhoff S, Casanovas A, Tillmann HL, Haro I (2011) Chem Biol Drug Des 78:277–282CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2012

Authors and Affiliations

  • Leticia Fernández
    • 1
  • M. José Bleda
    • 1
  • M. José Gómara
    • 1
  • Isabel Haro
    • 1
    Email author
  1. 1.Unit of Synthesis and Biomedical Applications of PeptidesIQAC-CSICBarcelonaSpain

Personalised recommendations