Abstract
Parvalbumin (PV) is a major fish allergen that is characterized by high water solubility, low molecular weight (i.e., ~12,000), acidic isoelectric point, heat stability and high phenylalanine content (compared with tyrosine and tryptophan). We used a fast, simple and efficient method to purify PV from <em>Cyprinus carpio</em> (carp). Heating, two runs of trichloroacetic acid precipitation and gel filtration chromatography were integrated in this method. Results of sodium dodecyl sulfate polyacrylamide gel electrophoresis and western blot showed that the purified protein was PV and the purity of carp PV obtained using this method was more than 90%. Moreover, the PV was characterized by MALDI-TOF-TOF MS and an unusually high 260 to 280 nm run absorbance ratio in the final purification stages. The rapid purification method used in this study resulted in better purification and identification of PV compared with other methods. Further studies on PV detection in freshwater fish can be performed based on the results of this experiment.
Highlights
The importance of fish as a food source has increased because of its high consumption and dietary value
Fish allergy in children and young adults is an immunoglobulin E-mediated disease with clinical manifestations ranging from urticaria and angioedema to anaphylaxis (Aas, 1966; Hansen et al, 1997)
The protein in lane 5 was greatly enriched after the second Trichloroacetic Acid (TCA) precipitation
Summary
The importance of fish as a food source has increased because of its high consumption and dietary value. Sampson (1999) found that the prevalence of fish and shellfish allergy in the United States is 0.5% of the total number of patients that show hypersensitivity to food items. Aas (1987) reported that 1/1000 of the general population in Norway is hypersensitive to fish. This value corresponds to approximately 2% of the adult population. Studies on various fish species including Cyprinus carpio (carp) (Bugajska-Schretter et al, 1999; Swoboda et al, 2002) salmon (Lindstrøm et al, 1996), cod
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