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Spot Synthesis: An Optimized Microarray to Detect IgE Epitopes

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Peptide Microarrays

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1352))

Abstract

Peptide microarrays have become increasingly more affordable in recent years with the SPOT technique being one of the most frequently used methods for synthesis and screening of peptides in arrays. Here, a protocol is presented for the identification of the amino acid sites involved in the conversion of human IgG to IgE response during the passive administration of therapeutic, anti-snake venom sera. Similarly, the minimal region of both the IgG and IgE binding epitopes, important for its interaction with ligand, were identified. As the ratio of concentrations for IgG to IgE in human serum is 1:10,000, also presented is a reproductive protocol of chemiluminescence-scanning for the detection of both immunoglobulins.

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Acknowledgement

This work received financial assistance from the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) and Fundação Carlos Chagas Filho de Amparo a Pesquisa do Estado do Rio de Janeiro (FAPERJ).

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Authors and Affiliations

  1. FIOCRUZ, Center of Technological Development in Health (CDTS) / National Institute of Science and Technology for Innovation on Neglected Diseases (INCT-IDN), FIOCRUZ, Rio de Janeiro, Brazil

    Salvatore G. De-Simone

  2. Department of Cellular andMolecularBiology, Federal FluminenseUniversity,Biology Institute, Niterói, Rio de Janeiro, Brazil

    Paloma Napoleão-Pêgo

  3. IOCRUZ, Center of Technological Development in Health (CDTS) / National Institute of Science and Technology for Innovation on Neglected Diseases (INCT-IDN) FIOCRUZ, Rio de Janeiro, Brazil

    Thatiane S. De-Simone

Authors
  1. Salvatore G. De-Simone
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  2. Paloma Napoleão-Pêgo
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  3. Thatiane S. De-Simone
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Corresponding author

Correspondence to Salvatore G. De-Simone .

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Editors and Affiliations

  1. Consiglio Nazionale delle Ricerche, Milano, Italy

    Marina Cretich

  2. Consiglio Nazionale delle Ricerche, Milano, Italy

    Marcella Chiari

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De-Simone, S.G., Napoleão-Pêgo, P., De-Simone, T.S. (2016). Spot Synthesis: An Optimized Microarray to Detect IgE Epitopes. In: Cretich, M., Chiari, M. (eds) Peptide Microarrays. Methods in Molecular Biology, vol 1352. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3037-1_20

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  • DOI: https://doi.org/10.1007/978-1-4939-3037-1_20

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3036-4

  • Online ISBN: 978-1-4939-3037-1

  • eBook Packages: Springer Protocols

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