LC-MS/MS Reveals Extraction Effects on Shellfish Allergens

Shellfish is one of the top causes of severe, life-threatening allergic reactions to food. When it comes to detecting and measuring the proteins that trigger these reactions (allergens), how the proteins are extracted from the shellfish in the first place plays a big role in the results. To dig into this, researchers looked at two commonly eaten shrimp species (black tiger prawn and white leg prawn) to see how different protein-extraction methods affected allergen detection. They used sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE), which separates proteins by size, along with a more advanced method called label-free liquid chromatography-tandem mass spectrometry (LC-MS/MS), to measure how much of each protein was present. Their findings were published in the International Journal of Biological Macromolecules.¹

What are Food Allergies, How Common Are They, and Why Are Shellfish (Particularly Prawns) a Significant Concern?

A food allergy occurs when the immune system overreacts to certain substances in food (usually proteins) after being exposed to them.^2,3 Roughly 1 to 10% of people worldwide are affected by food allergies, and that number has been climbing in recent decades.⁴Shellfish and fish, two of the nine foods that account for most food allergies, are notorious for allergies that people rarely outgrow, and together they're one of the leading causes of severe, life-threatening allergic reactions.^5,6 Shellfish include two main types: crustaceans (like shrimp and crab) and mollusks (like clams and mussels). Among crustaceans, prawns are eaten the most and are a leading trigger of shellfish allergies.⁷

How Did the Extraction Method Affect Shrimp Allergen Detection, and What Allergens Did the Analysis Uncover?

The researchers pulled proteins from both raw and cooked shrimp muscle using four different extraction solutions: urea-based, sodium dodecyl sulphate (SDS)-based, SDS-based with reducing agent, and phosphate-buffered saline (PBS). They then used the computer program AllerCatPro to predict which proteins were likely to trigger allergies and followed up by testing how strongly those proteins reacted with allergy antibodies.¹

It turned out that the extraction method mattered a lot; it changed how many proteins were detected overall, how strongly they reacted with antibodies, how many different allergens showed up, and how much of each allergen was present. The PBS solution, despite being the standard choice, pulled out the highest overall number of allergens (roughly 84-95%) but actually captured the fewest different types of allergens, and it underrepresented some important ones, like tropomyosin and myosin light chain in raw shrimp, and arginine kinase in cooked shrimp. On the other hand, the more aggressive extraction solutions that break down protein structure captured a wider range of allergens, with the urea-based solution used on cooked shrimp turning up the most complete allergen picture.¹

Beyond the allergens already well known to science, the researchers also found up to 72 additional low-level proteins that computer analysis strongly suggested could be allergenic, and how much of each one showed up depended heavily on which extraction method was used. Overall, the findings suggest that fine-tuning the protein-extraction process can lead to better allergen detection, helping uncover overlooked or hard-to-detect allergens that deserve further testing, and ultimately supporting more thorough allergy risk assessments.¹

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References

1. Iddagoda, J.; Chin, R.; Karnaneedi, S. et al. Protein Extraction Strategies Shape Shellfish Allergen Landscapes: Implications for Seafood Safety, Allergy Diagnosis and Risk Management. Int J Biol Macromol. 2026, 153882. DOI: 10.1016/j.ijbiomac.2026.153882
2. Renz, H.; Allen, K. J.; Sicherer, S. H. et al. Food Allergy. Nat. Rev. Dis.Primers 2018, 4. DOI:10.1038/nrdp.2017.98
3. Yu, W.; Freeland, D. M. H.; Nadeau, K. C. Food Allergy: Immune Mechanisms, Diagnosis and Immunotherapy. Nat. Rev. Immunol. 2016, 16, 751-765. DOI: 10.1038/nri.2016.111
4. Leung, A. S. Y.; Xing, Y.; Fernández-Rivas, M. et al. The Relationship Between Dietary Patterns and the Epidemiology of Food Allergy. Allergy2025, 80, 690-702. DOI: 10.1111/all.16455
5. Zotova, V.; Clarke, A. E.; Chan, E. S. et al. Low Resolution Rates of Seafood Allergy. J Allergy Clin Immunol Pract.2019, 7, 690-692. DOI: 10.1016/j.jaip.2018.09.011
6. Davis, C. M.; Gupta, R. S.; Aktas, O. N. et al. Clinical Management of Seafood Allergy. J Allergy Clin Immunol Pract.2020, 8, 37-44. DOI: 10.1016/j.jaip.2019.10.019
7. Chen, J.; Zhang, Q.; Ying, Y. et al. Prevalence of Shrimp Allergy: A Meta-Analysis Based on Different Diagnostic Methods. Front. Allergy2025, 6. DOI: 10.3389/falgy.2025.1635274