Abstract
| Original language | English |
|---|---|
| Pages (from-to) | 271-281 |
| Number of pages | 11 |
| Journal | Journal of Immunological Methods |
| Volume | 278 |
| Issue number | 1-2 |
| DOIs | |
| Publication status | Published - 2003 |
| MoE publication type | A1 Journal article-refereed |
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Keywords
- Allergen
- Allergy
- Hevein
- IgE antibody
- Latex
- Phage display
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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 journal › Article › Scientific › peer-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 -