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


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.


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



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)


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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

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