Development of a mass spectrometry immunoassay for unambiguous detection of egg allergen traces in wines

A mass spectrometry immunoassay (MSIA) specifically designed for the detection of egg allergens in wines is described. MSIA is based on an immunoaffinity enrichment procedure combined with targeted MS/MS detection of selected egg peptide markers. Polyclonal antibodies raised against native ovalbumin, chosen as the target protein tracing for egg powder, were immobilized onto low backpressure monolithic MSIA customized disposable tips. Ovalbumin-free wine samples were fortified with standard protein at different concentrations in the low microgram-per-milliliter range. A simple protocol was devised consisting of a 1:4 dilution of the wine sample with a basic solution for pH adjustment, followed by a semi-automated purification/enrichment step on MSIA customized disposable tips fitted on a multichannel electronic pipette. Among the main figures of merit, LOD and LOQ values as low as 0.01 and 0.03 μg/mL, respectively, and within-day precision of 18% should be noticed. Noteworthy, the developed assay outperformed current MS-based methods for the detection of allergenic protein in wine matrices, thanks to the immunoaffinity enrichment. In addition, compared to other immunoassays, the present approach boasts the unquestionable advantage of providing an unambiguous identification of the target protein by simultaneous detection of three unique peptide markers each giving three specific MS/MS transitions.

[1]  Rosa Pilolli,et al.  Chapter 7 - Mass Spectrometry in Food Allergen Research , 2015 .

[2]  P. Restani,et al.  Collaborative Interlaboratory Studies for the Validation of ELISA Methods for the Detection of Allergenic Fining Agents Used in Wine According to the Criteria of OIV Resolution 427–2010 Modified by OIV–Comex 502–2012 , 2014, Food Analytical Methods.

[3]  P. Righetti,et al.  Proteomics of wine additives: mining for the invisible via combinatorial peptide ligand libraries. , 2010, Journal of proteomics.

[4]  A. Prakash,et al.  An automated, high‐throughput method for targeted quantification of intact insulin and its therapeutic analogs in human serum or plasma coupling mass spectrometric immunoassay with high resolution and accurate mass detection (MSIA‐HR/AM) , 2014, Proteomics.

[5]  P. Restani,et al.  Immunochemical investigation of allergenic residues in experimental and commercially-available wines fined with egg white proteins. , 2014, Food chemistry.

[6]  EFSA Panel on Dietetic Products, Nutrition and Allergies (NDA); Scientific Opinion related to a notification from the International Organisation of Vine and Wine (OIV) on casein/caseinate/milk products to be used in the manufacture of wine as clarification , 2011 .

[7]  Rosa Pilolli,et al.  Multi-allergen quantification of fining-related egg and milk proteins in white wines by high-resolution mass spectrometry. , 2013, Rapid communications in mass spectrometry : RCM.

[8]  M. Thevis,et al.  Qualitative identification of growth hormone-releasing hormones in human plasma by means of immunoaffinity purification and LC-HRMS/MS , 2016, Analytical and Bioanalytical Chemistry.

[9]  M. Fischer,et al.  Impact of wine manufacturing practice on the occurrence of fining agents with allergenic potential , 2014, Food additives & contaminants. Part A, Chemistry, analysis, control, exposure & risk assessment.

[10]  R. Pilolli,et al.  Challenging the Limit of Detection for Egg Allergen Detection in Red Wines by Surface Plasmon Resonance Biosensor , 2016, Food Analytical Methods.

[11]  Robin Gerlach,et al.  CORRIGENDUM: Taxis Toward Hydrogen Gas by Methanococcus maripaludis , 2014, Scientific Reports.

[12]  P. Righetti,et al.  Noah's nectar: the proteome content of a glass of red wine. , 2010, Journal of proteomics.

[13]  Randall W. Nelson,et al.  Parallel Workflow for High-Throughput (>1,000 Samples/Day) Quantitative Analysis of Human Insulin-Like Growth Factor 1 Using Mass Spectrometric Immunoassay , 2014, PloS one.

[14]  F. Regnier,et al.  Mass-linked immuno-selective assays in targeted proteomics. , 2013, Analytical chemistry.

[15]  Mary F. Lopez,et al.  A semi-automated mass spectrometric immunoassay coupled to selected reaction monitoring (MSIA-SRM) reveals novel relationships between circulating PCSK9 and metabolic phenotypes in patient cohorts. , 2015, Methods.

[16]  Jennifer N. Sutton,et al.  Rapid development of sensitive, high-throughput, quantitative and highly selective mass spectrometric targeted immunoassays for clinically important proteins in human plasma and serum. , 2013, Clinical biochemistry.

[17]  R. Pilolli,et al.  Coupling SPE on-line pre-enrichment with HPLC and MS/MS for the sensitive detection of multiple allergens in wine , 2017 .

[18]  Randall W. Nelson,et al.  Mass spectrometric immunoassay , 1996, Photonics West.

[19]  D. Billheimer,et al.  Mass spectrometric immunoassay and MRM as targeted MS‐based quantitative approaches in biomarker development: Potential applications to cardiovascular disease and diabetes , 2013, Proteomics. Clinical applications.

[20]  H. Nakagawa,et al.  Antibody-coupled monolithic silica microtips for highthroughput molecular profiling of circulating exosomes , 2014, Scientific Reports.

[21]  Ilario Losito,et al.  Development of a method for the quantification of caseinate traces in Italian commercial white wines based on liquid chromatography-electrospray ionization-ion trap-mass spectrometry. , 2013, Journal of agricultural and food chemistry.

[22]  U. Immer,et al.  Collaborative Tests of ELISA Methods for the Determination of Egg White Protein and Caseins Used as Fining Agents in Red and White Wines , 2014, Food Analytical Methods.

[23]  Angelo Visconti,et al.  Immunochemical and DNA-based methods in food allergen analysis and quality assurance perspectives. , 2010 .

[24]  Linda Monaci,et al.  Mass spectrometry-based proteomics methods for analysis of food allergens , 2009 .

[25]  G. Arrigoni,et al.  Analysis of commercial wines by LC-MS/MS reveals the presence of residual milk and egg white allergens , 2012 .

[26]  B. Domon,et al.  Screening protein isoforms predictive for cancer using immunoaffinity capture and fast LC‐MS in PRM mode , 2015, Proteomics. Clinical applications.

[27]  Antonio Masi,et al.  Mass spectrometry detection of egg proteins in red wines treated with egg white , 2012 .

[28]  L. Jacxsens,et al.  Analysis to support allergen risk management: Which way to go? , 2013, Journal of agricultural and food chemistry.

[29]  Maria Careri,et al.  Investigation of different sample pre-treatment routes for liquid chromatography–tandem mass spectrometry detection of caseins and ovalbumin in fortified red wine , 2014 .

[30]  N. Bache,et al.  Detection and differentiation of 22 kDa and 20 kDa Growth Hormone proteoforms in human plasma by LC-MS/MS. , 2015, Biochimica et biophysica acta.

[31]  Rosa Pilolli,et al.  Rapid and label-free detection of egg allergen traces in wines by surface plasmon resonance biosensor , 2015, Analytical and Bioanalytical Chemistry.

[32]  Rosa Pilolli,et al.  Orbitrap™ monostage MS versus hybrid linear ion trap MS: application to multi-allergen screening in wine. , 2014, Journal of mass spectrometry : JMS.

[33]  Francesca Uberti,et al.  Allergenic Proteins in Enology: A Review on Technological Applications and Safety Aspects , 2015, Molecules.

[34]  R. Nelson,et al.  Development of multiplex mass spectrometric immunoassay for detection and quantification of apolipoproteins C-I, C-II, C-III and their proteoforms. , 2015, Methods.