Hevein-specific recombinant IgE antibodies from human single-chain antibody phage display libraries

Marja-Leena Laukkanen, Soili Mäkinen-Kiljunen, Kirsi Isoherranen, Tari Haahtela, Hans Söderlund, Kristiina Takkinen

Research output: Contribution to journalArticleScientificpeer-review

23 Citations (Scopus)

Abstract

IgE antibodies distinctively recognising allergenic epitopes would be ideal reagents in immunodiagnostics to detect and quantify allergens, as well as for the development of allergy diagnostics and therapeutics. We have isolated recombinant human IgE antibodies specific for the major latex allergen, hevein, from antibody phage display libraries using a green fluorescent protein (GFP)–hevein fusion as a selection antigen. Human IgE phage display libraries were constructed by combining the IgE heavy chain genes to kappa and lambda light-chain genes which were isolated from lymphocytes of a latex allergic patient. The screening of antibody libraries resulted in the enrichment of two hevein-binding scFvs designated as 1A4 and 1C2. Both antibodies showed specific binding to the hevein that could be inhibited by both the recombinant GFP–hevein and native hevein isolated from latex examination gloves. The scFvs were prone to aggregate and, thus, for further characterisation, they were converted to Fab fragments with human IgG1 or IgE isotype. Similar hevein-binding properties of the 1A4 and 1C2 Fab fragments and human IgE serum pool, conventionally used in the detection of latex allergens, demonstrate the potential utility of these recombinant antibodies for the analysis of latex allergen.
Original languageEnglish
Pages (from-to)271-281
Number of pages11
JournalJournal of Immunological Methods
Volume278
Issue number1-2
DOIs
Publication statusPublished - 2003
MoE publication typeA1 Journal article-refereed

Keywords

  • Allergen
  • Allergy
  • Hevein
  • IgE antibody
  • Latex
  • Phage display

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