Differentiating cross-reacting allergens in the immunological analysis of celery (Apium graveolens) by mass spectrometry.
Celery is acknowledged as a major food allergen in Europe, and mandatory labeling for preprocessed foods has been implemented. However, no methods for the specific detection of celery protein in foods have been published. In the present study, a sandwich celery ELISA using polyclonal anticelery antibodies for capture and detection was developed and validated. The method has an LOD of 0.5 mg/kg in buffer; however, it is applicable only for the screening of food products because of extensive cross-reactivity with potato and carrot proteins. Using nanoLC-ion-trap MS/MS, a number of proteins in the three vegetable species were identified as candidates for causing cross-reactions due to amino acid sequence homologies. Among others, a novel patatin (Sola t 1)-like protein was detected in celery and a flavin adenine dinucleotide binding domain-containing protein (Api g 5)-like protein was identified in carrot. The utility of triple-quadrupole MS/MS for specific and quantitative analysis of celery, potato, and carrot allergens was evaluated using whole protein extracts. Several unique precursor ion-to-product ion transitions were determined for each species, suggesting the feasibility of developing an MS-based screening method to specifically detect celery allergens in foods.
Faeste
Christiane K
CK
National Veterinary Institute, Department of Feed and Food Safety, Section of Chemistry, PO Box 750 Sentrum, 0106 Oslo, Norway. christiane.faste@vetinst.no
Jonscher
Karen R
KR
Sit
Louis
L
Klawitter
Jelena
J
Løvberg
Kjersti E
KE
Moen
Lena H
LH
D000485:Allergens / Q000737:chemistry / Q000276:immunology*; D000595:Amino Acid Sequence; D052179:Antigens, Plant; D000077102:Apium; D002021:Buffers; D002265:Carboxylic Ester Hydrolases / Q000737:chemistry; D003429:Cross Reactions / Q000276:immunology*; D018552:Daucus carota; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005512:Food Hypersensitivity / Q000276:immunology*; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods*; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000737:chemistry; D015203:Reproducibility of Results; D017386:Sequence Homology, Amino Acid; D053719:Tandem Mass Spectrometry / Q000379:methods*; D014675:Vegetables
D000485:Allergens; D052179:Antigens, Plant; C477756:Api g 5 allergen, celery; D002021:Buffers; D010940:Plant Proteins; C043222:patatin protein, Solanum tuberosum; D002265:Carboxylic Ester Hydrolases