Selection of recombinant IgE antibodies binding the ß-lactoglobulin allergen in a conformation-dependent manner

Sirpa Jylhä, Soili Mäkinen-Kiljunen, Tari Haahtela, Hans Söderlund, Kristiina Takkinen, Marja-Leena Laukkanen (Corresponding Author)

Research output: Contribution to journalArticleScientificpeer-review

11 Citations (Scopus)

Abstract

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 β-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.
Original languageEnglish
Pages (from-to)63-70
Number of pages8
JournalJournal of Immunological Methods
Volume350
Issue number1-2
DOIs
Publication statusPublished - 2009
MoE publication typeA1 Journal article-refereed

Fingerprint

Lactoglobulins
Allergens
Immunoglobulin E
Antibodies
Milk
Immunoglobulin Fab Fragments
Clone Cells
Milk Hypersensitivity
Food Hypersensitivity
Libraries
Hypersensitivity
Immunoglobulin G
Hot Temperature
Food

Keywords

  • Allergen
  • Allergen detection
  • beta-lactoglobulin
  • Cow's milk
  • IgE antibody
  • Phage display

Cite this

Jylhä, Sirpa ; Mäkinen-Kiljunen, Soili ; Haahtela, Tari ; Söderlund, Hans ; Takkinen, Kristiina ; Laukkanen, Marja-Leena. / Selection of recombinant IgE antibodies binding the ß-lactoglobulin allergen in a conformation-dependent manner. In: Journal of Immunological Methods. 2009 ; Vol. 350, No. 1-2. pp. 63-70.
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abstract = "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 β-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.",
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Selection of recombinant IgE antibodies binding the ß-lactoglobulin allergen in a conformation-dependent manner. / Jylhä, Sirpa; Mäkinen-Kiljunen, Soili; Haahtela, Tari; Söderlund, Hans; Takkinen, Kristiina; Laukkanen, Marja-Leena (Corresponding Author).

In: Journal of Immunological Methods, Vol. 350, No. 1-2, 2009, p. 63-70.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Selection of recombinant IgE antibodies binding the ß-lactoglobulin allergen in a conformation-dependent manner

AU - Jylhä, Sirpa

AU - Mäkinen-Kiljunen, Soili

AU - Haahtela, Tari

AU - Söderlund, Hans

AU - Takkinen, Kristiina

AU - Laukkanen, Marja-Leena

PY - 2009

Y1 - 2009

N2 - 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 β-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.

AB - 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 β-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.

KW - Allergen

KW - Allergen detection

KW - beta-lactoglobulin

KW - Cow's milk

KW - IgE antibody

KW - Phage display

U2 - 10.1016/j.jim.2009.07.007

DO - 10.1016/j.jim.2009.07.007

M3 - Article

VL - 350

SP - 63

EP - 70

JO - Journal of Immunological Methods

JF - Journal of Immunological Methods

SN - 0022-1759

IS - 1-2

ER -