Selection of recombinant IgE antibodies binding the beta-lactoglobulin allergen in a conformation-dependent manner.
Cow's milk allergy (CMA) is a common food allergy, especially among infants and young children. Approximately 85% of milk-allergic children outgrow their allergy by the age of three but the remaining 15% remain allergic. Bovine beta-lactoglobulin (BLG) is one of the major allergens in cow's milk. There is a definite need for the specific and sensitive detection of allergenic substances. Validated methods are obligatory to demonstrate allergen contamination and even fatal hidden allergens and, thus, to prevent life-threatening conditions of allergic persons. In this study, we constructed human IgE scFv libraries from an adult milk-allergic patient and isolated the first recombinant IgE antibodies specific to a food allergen, BLG. The selection of the IgE antibody libraries with two distinct panning procedures resulted in the enrichment of four clones having different BLG-binding profiles; two of the clones recognize the native BLG whereas the other two recognize only the heat-denatured form of BLG. For further characterization, the scFv fragments were converted to Fab fragments with human IgG1 isotype. The D1 Fab fragment, binding native BLG with nanomolar affinity, also partially inhibited serum IgE binding to BLG. These BLG-specific IgE antibodies can be applied for the detection of both native and denatured BLG in cow's milk products and furthermore, for the optimization of manufacturing processes to develop safe hypoallergenic milk products.
Jylhä
Sirpa
S
VTT Technical Research Centre of Finland, P.O. Box 1000, FIN-02044 VTT, Finland.
Mäkinen-Kiljunen
Soili
S
Haahtela
Tari
T
Söderlund
Hans
H
Takkinen
Kristiina
K
Laukkanen
Marja-Leena
ML
D000328:Adult; D000485:Allergens / Q000032:analysis / Q000737:chemistry* / Q000276:immunology; D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000235:genetics / Q000276:immunology; D002417:Cattle; D002648:Child; D002675:Child, Preschool; D003001:Cloning, Molecular / Q000379:methods*; D005504:Food Analysis / Q000379:methods; D015723:Gene Library; D006801:Humans; D007073:Immunoglobulin E / Q000737:chemistry* / Q000235:genetics / Q000276:immunology; D007782:Lactoglobulins / Q000032:analysis / Q000737:chemistry* / Q000276:immunology; D016269:Milk Hypersensitivity / Q000276:immunology / Q000517:prevention & control; D011487:Protein Conformation; D011489:Protein Denaturation; D017510:Protein Folding
D000485:Allergens; D000911:Antibodies, Monoclonal; D007782:Lactoglobulins; D007073:Immunoglobulin E