Monoclonal antibody specific to a major fish allergen: parvalbumin.
The major fish allergen, parvalbumin, is a low-molecular-weight (10 to 13 kDa), heat-stable protein. Monoclonal antibody (MAb) 3E1, developed against heat-treated catfish sarcoplasmic protein extract, recognizes a thermal-stable protein with the molecular-weight range of parvalbumin in fish extracts. We further investigated the antigen-binding characteristics of this antibody by comparing its immunoreactivity against various fish and other animal species, with a commercially available anti-parvalbumin antibody, MAb PARV-19. Soluble proteins were extracted from 67 cooked (100 degrees C for 20 min) finfish, shellfish, meat, and poultry species. Indirect enzyme-linked immunosorbent assay (ELISA) was performed to examine the immunoreactivity of both MAb 3E1 and MAb PARV-19 with sample extracts. Western blot was performed to compare the antigenic protein banding patterns in cooked fish extracts by using these two MAbs. The ELISA results revealed that both MAbs had identical reaction patterns to the fish species tested. Removal of Ca2+ from the fish extracts increased the overall immunoreactivity of both MAbs. Western blot results confirmed that the antigenic protein banding pattern in various fish species blotted by MAb 3E1 corresponded to the molecular weights of parvalbumins recognized by PARV-19. However, screening with non-finfish extracts revealed MAb 3E1 to be strictly finfish specific, while PARV-19 cross-reacted with frog, rat, and rabbit extracts. Based on the heat stability, molecular weight, immunoreactivity, and Ca2+-dependent binding of the antigenic proteins, MAb 3E1 is specific to fish parvalbumin. It would therefore be a useful probe for investigating the major fish allergen in both raw and processed food.
Gajewski
Kamil G
KG
Department of Nutrition, Food and Exercise Sciences, 420 Sandels Building, Florida State University, Tallahassee, Florida 32306-1493, USA.
Hsieh
Yun-Hwa P
YH
D000485:Allergens / Q000737:chemistry / Q000276:immunology*; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000941:Antigens / Q000276:immunology; D001001:Anura; D002118:Calcium / Q000737:chemistry; D000939:Epitopes; D005260:Female; D005399:Fishes; D051379:Mice; D008807:Mice, Inbred BALB C; D010320:Parvalbumins / Q000276:immunology*; D013045:Species Specificity
D000485:Allergens; D000911:Antibodies, Monoclonal; D000941:Antigens; D000939:Epitopes; D010320:Parvalbumins; D002118:Calcium