“…The major allergen in shrimp, TM, was specifically targeted for purification using multiple precipitation steps and then introduced to a recently developed protocol for primary structure determination. , This protocol is based on using multiple enzymatic digestions, different MS ion sources, and a derivatization reaction to sequence the global protein with 100% amino acid coverage as shown in Figure . Three enzymes were used, trypsin, Gul-C V8, and ASP-N, to increase the diversity of the produced peptides using two MS ion sources, ESI and MALDI.…”
Section: Resultsmentioning
confidence: 99%
“…While occupational respiratory diseases are still largely under-recognized, they remain poorly diagnosed and managed. − Various epidemiological studies among seafood processors indicate that the prevalence of occupational asthma is between 2% and 36%, while it is more commonly associated with shellfish processing . The challenge for bioaerosol exposure assessment is the lack of methodological advancements in the accurate and sensitive quantification of biomarker exposure .…”
Section: Discussionmentioning
confidence: 99%
“…This study introduces a functional proteomics strategy (allergenomic) to evaluate the potential allergenic proteins in northern shrimp (NS) as summarized in Figure . Sera from sensitized patients were used to evaluate the proteomics profile of NS which was subsequently identified using mass spectrometry.…”
Occupational asthma is a major chronic health dilemma among workers involved in the seafood industry. Several proteins notoriously known to cause asthma have been reported in different seafood. This work involves the application of an allergenomics strategy to study the most potent allergens of northern shrimp. The proteins were extracted from shrimp tissue and profiled by gel electrophoresis. Allergenic proteins were identified based on their reactivity to patient sera and were structurally identified using tandem mass spectrometry. Northern shrimp tropomyosin, arginine kinase, and sarcoplasmic calcium-binding protein were found to be the most significant allergens. Multiple proteolytic enzymes enabled 100% coverage of the sequence of shrimp tropomyosin by tandem mass specrometry. Only partial sequence coverage was obtained, however, for the shrimp allergen arginine kinase. Signature peptides, for both tropomyosin and arginine kinase, were assigned and synthesized for use in developing the multiple reaction monitoring tandem mass spectrometric method. Subsequently, air samples were collected from a shrimp processing plant and two aerosolized proteins quantified using tandem mass specrometry. Allergens were detected in all areas of the plant, reaching levels as high as 375 and 480 ng/m(3) for tropomyosine and arginine kinase, respectively. Tropomyosine is much more abundant than arginine kinase in shrimp tissues, so the high levels of arginine kinase suggest it is more easily aerosolized. The present study shows that mass spectrometric analysis is a sensitive and accurate tool in identifying and quantifying aerosolized allergens.
“…The major allergen in shrimp, TM, was specifically targeted for purification using multiple precipitation steps and then introduced to a recently developed protocol for primary structure determination. , This protocol is based on using multiple enzymatic digestions, different MS ion sources, and a derivatization reaction to sequence the global protein with 100% amino acid coverage as shown in Figure . Three enzymes were used, trypsin, Gul-C V8, and ASP-N, to increase the diversity of the produced peptides using two MS ion sources, ESI and MALDI.…”
Section: Resultsmentioning
confidence: 99%
“…While occupational respiratory diseases are still largely under-recognized, they remain poorly diagnosed and managed. − Various epidemiological studies among seafood processors indicate that the prevalence of occupational asthma is between 2% and 36%, while it is more commonly associated with shellfish processing . The challenge for bioaerosol exposure assessment is the lack of methodological advancements in the accurate and sensitive quantification of biomarker exposure .…”
Section: Discussionmentioning
confidence: 99%
“…This study introduces a functional proteomics strategy (allergenomic) to evaluate the potential allergenic proteins in northern shrimp (NS) as summarized in Figure . Sera from sensitized patients were used to evaluate the proteomics profile of NS which was subsequently identified using mass spectrometry.…”
Occupational asthma is a major chronic health dilemma among workers involved in the seafood industry. Several proteins notoriously known to cause asthma have been reported in different seafood. This work involves the application of an allergenomics strategy to study the most potent allergens of northern shrimp. The proteins were extracted from shrimp tissue and profiled by gel electrophoresis. Allergenic proteins were identified based on their reactivity to patient sera and were structurally identified using tandem mass spectrometry. Northern shrimp tropomyosin, arginine kinase, and sarcoplasmic calcium-binding protein were found to be the most significant allergens. Multiple proteolytic enzymes enabled 100% coverage of the sequence of shrimp tropomyosin by tandem mass specrometry. Only partial sequence coverage was obtained, however, for the shrimp allergen arginine kinase. Signature peptides, for both tropomyosin and arginine kinase, were assigned and synthesized for use in developing the multiple reaction monitoring tandem mass spectrometric method. Subsequently, air samples were collected from a shrimp processing plant and two aerosolized proteins quantified using tandem mass specrometry. Allergens were detected in all areas of the plant, reaching levels as high as 375 and 480 ng/m(3) for tropomyosine and arginine kinase, respectively. Tropomyosine is much more abundant than arginine kinase in shrimp tissues, so the high levels of arginine kinase suggest it is more easily aerosolized. The present study shows that mass spectrometric analysis is a sensitive and accurate tool in identifying and quantifying aerosolized allergens.
“…Eight different crustacean allergens have been investigated by three different research groups (Table and Supplementary Table 1 in the Supporting Information). ,− Arginine kinase is the most investigated allergen in this allergen source. Ortea et al attempted to identify different peptides for different species with MALDI and IT.…”
Section: Advantages Of Mass Spectrometry Systems For
Allergen Detectionmentioning
Food allergies are increasing worldwide and becoming a public health concern. Food legislation requires detailed declarations of potential allergens in food products and therefore an increased capability to analyze for the presence of food allergens. Currently, antibody-based methods are mainly utilized to quantify allergens; however, these methods have several disadvantages. Recently, mass spectrometry (MS) techniques have been developed and applied to food allergen analysis. At present, 46 allergens from 11 different food sources have been characterized using different MS approaches and some specific signature peptides have been published. However, quantification of allergens using MS is not routinely employed. This review compares the different aspects of food allergen quantification using advanced MS techniques including multiple reaction monitoring. The latter provides low limits of quantification for multiple allergens in simple or complex food matrices, while being robust and reproducible. This review provides an overview of current approaches to analyze food allergens, with specific focus on MS systems and applications.
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