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

Fingerprint

Single-Chain Antibodies
Bacteriophages
Immunoglobulin E
Latex
Libraries
Allergens
Antibodies
Immunoglobulin Fab Fragments
Green Fluorescent Proteins
Genes
hevein
Epitopes
Hypersensitivity
Immunoglobulin G
Lymphocytes
Light
Antigens
Serum

Keywords

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

Cite this

Laukkanen, Marja-Leena ; Mäkinen-Kiljunen, Soili ; Isoherranen, Kirsi ; Haahtela, Tari ; Söderlund, Hans ; Takkinen, Kristiina. / Hevein-specific recombinant IgE antibodies from human single-chain antibody phage display libraries. In: Journal of Immunological Methods. 2003 ; Vol. 278, No. 1-2. pp. 271-281.
@article{d9563ae6e775488295bd81a7c5695d11,
title = "Hevein-specific recombinant IgE antibodies from human single-chain antibody phage display libraries",
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.",
keywords = "Allergen, Allergy, Hevein, IgE antibody, Latex, Phage display",
author = "Marja-Leena Laukkanen and Soili M{\"a}kinen-Kiljunen and Kirsi Isoherranen and Tari Haahtela and Hans S{\"o}derlund and Kristiina Takkinen",
year = "2003",
doi = "10.1016/S0022-1759(03)00070-X",
language = "English",
volume = "278",
pages = "271--281",
journal = "Journal of Immunological Methods",
issn = "0022-1759",
publisher = "Elsevier",
number = "1-2",

}

Hevein-specific recombinant IgE antibodies from human single-chain antibody phage display libraries. / Laukkanen, Marja-Leena; Mäkinen-Kiljunen, Soili; Isoherranen, Kirsi; Haahtela, Tari; Söderlund, Hans; Takkinen, Kristiina.

In: Journal of Immunological Methods, Vol. 278, No. 1-2, 2003, p. 271-281.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Hevein-specific recombinant IgE antibodies from human single-chain antibody phage display libraries

AU - Laukkanen, Marja-Leena

AU - Mäkinen-Kiljunen, Soili

AU - Isoherranen, Kirsi

AU - Haahtela, Tari

AU - Söderlund, Hans

AU - Takkinen, Kristiina

PY - 2003

Y1 - 2003

N2 - 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.

AB - 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.

KW - Allergen

KW - Allergy

KW - Hevein

KW - IgE antibody

KW - Latex

KW - Phage display

U2 - 10.1016/S0022-1759(03)00070-X

DO - 10.1016/S0022-1759(03)00070-X

M3 - Article

VL - 278

SP - 271

EP - 281

JO - Journal of Immunological Methods

JF - Journal of Immunological Methods

SN - 0022-1759

IS - 1-2

ER -