Purification and characterization of natural Bet v 1 from birch pollen and related allergens from carrot and celery

Autor: Mirko A. Bollen, Aranzazu Garcia, Johannes P. F. G. Helsper, Harry J. Wichers, Martinus A.J.S. van Boekel, Jan H.G. Cordewener, Huub F. J. Savelkoul
Jazyk: angličtina
Rok vydání: 2007
Předmět:
Circular dichroism
cross-reactivity
medicine.disease_cause
Cross-reactivity
Protein Structure
Secondary
law.invention
molecular characterization
Allergen
Tandem Mass Spectrometry
law
Protein Isoforms
Trypsin
Betula
Plant Proteins
medicine.diagnostic_test
Chemistry
Circular Dichroism
Hydrophilic interaction chromatography
apium-graveolens
Product Design and Quality Management Group
Daucus carota
PRI Bioscience
Recombinant DNA
Pollen
Electrophoresis
Polyacrylamide Gel
apple allergen
pru av-1
Biotechnology
Blotting
Western
Molecular Sequence Data
Size-exclusion chromatography
Celbiologie en Immunologie
Western blot
medicine
Amino Acid Sequence
low-temperature method
Ammonium sulfate precipitation
Apium
VLAG
Chromatography
AFSG Food Quality
Leerstoelgroep Productontwerpen en kwaliteitskunde
bet-v-i
Allergens
Antigens
Plant
mass-spectrometry
Cell Biology and Immunology
WIAS
plant-tissues
Sequence Alignment
Food Science
major cherry allergen
Zdroj: Molecular Nutrition & Food Research 51 (2007) 12
Molecular Nutrition & Food Research, 51(12), 1527-1536
ISSN: 1613-4125
1527-1536
DOI: 10.1002/mnfr.200700119
Popis: Birch pollen allergy is predominantly caused by the major allergen Bet v 1 and can lead to crossreactions with homologous proteins in food. Two major cross-reactive food allergens are Dau c 1 from carrot and Api g 1 from celery, which have never been purified from their natural source. Here, we describe a non-denaturing purification method for obtaining natural Bet v 1, Dau c 1 and Api g 1, comprising of ammonium sulfate precipitation, hydrophobic interaction chromatography and size exclusion chromatography. This method resulted in 98-99% pure isoform mixtures for each allergen. Characterization of these isoform mixtures with Q-TOF MS/MS clearly showed earlier reported isoforms of Bet v 1, Dau c 1 and Api g 1, but also new isoforms. The presence of secondary structure in the three purified allergens was demonstrated via circular dichroism and showed high similarity. The immune reactivity of the natural allergens was compared with recombinant proteins by Western blot and ELISA and showed similar reactivity.
Databáze: OpenAIRE
Externí odkaz: