Hevein-specific recombinant IgE antibodies from human single-chain antibody phage display libraries.
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.
Laukkanen
Marja-Leena
ML
VTT Biotechnology, P.O. Box 1500, FIN-02044 VTT, Finland. marja-leena.laukkanen@vtt.fi
Mäkinen-Kiljunen
Soili
S
Isoherranen
Kirsi
K
Haahtela
Tari
T
Söderlund
Hans
H
Takkinen
Kristiina
K
D000595:Amino Acid Sequence; D000818:Animals; D000918:Antibody Specificity; D023181:Antimicrobial Cationic Peptides; D016367:Baculoviridae / Q000235:genetics; D001667:Binding, Competitive; D003001:Cloning, Molecular; D017931:DNA Primers; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes; D049452:Green Fluorescent Proteins; D006801:Humans; D015151:Immunoblotting; D007073:Immunoglobulin E / Q000737:chemistry / Q000302:isolation & purification*; D007140:Immunoglobulin Fab Fragments / Q000276:immunology; D007074:Immunoglobulin G / Q000737:chemistry / Q000302:isolation & purification; D007840:Latex / Q000737:chemistry / Q000276:immunology*; D020315:Latex Hypersensitivity / Q000175:diagnosis / Q000276:immunology*; D008164:Luminescent Proteins; D008969:Molecular Sequence Data; D019151:Peptide Library; D037121:Plant Lectins / Q000276:immunology*; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000276:immunology / Q000302:isolation & purification; D018411:Spodoptera / Q000821:virology
D023181:Antimicrobial Cationic Peptides; D017931:DNA Primers; D000939:Epitopes; D007140:Immunoglobulin Fab Fragments; D007074:Immunoglobulin G; D007840:Latex; D008164:Luminescent Proteins; D019151:Peptide Library; D037121:Plant Lectins; D011994:Recombinant Proteins; C045875:hevein; D049452:Green Fluorescent Proteins; D007073:Immunoglobulin E