Development and Characterization of Monoclonal Antibodies for the Detection of Fish Protein

Abstract

This study developed and characterized anti-fish monoclonal antibodies (mAbs) capable of detecting fish, a major allergenic food, in processed food products to protect fish sensitized individuals. Of the three mAbs raised against crude protein extract of cooked fish muscle, mAb 8F5 exhibited a positive reaction to all 50 common food fish species tested with no cross-reactions to shellfish, land animals, or food additives. Although the ELISA results were negative against swordfish and yellowfin tuna, western blot clearly detected both after cooking. The ~36 kDa antigenic protein of mAb 8F5, which was found in all fish species, was detectable by mAb 8F5 in all of the fish samples even after prolonged heat treatment (100 °C, up to 60 min). These findings suggest that mAb 8F5 has great potential utility as a probe for the immunochemical detection of fish tissue in cooked food.

Detection of a Thermal Stable-Soluble Protein (TSSP) as a Marker of Peanut Adulteration Using a Highly Sensitive Indirect Enzyme-Linked Immunosorbent Assay based on Monoclonal Antibodies

“…Thus, the RO 3A1-12 MAb reacted with different-size proteins at 22, 40, and 60 kDa, and PB 5F9-23 MAb bound to 32 and 64 kDa proteins. Chen and Hsieh (2021) also reported that monoclonal antibodies against TSSP in fish meat reacted simultaneously with proteins of different sizes and presumed that the protein was degraded or polymerized as we mentioned above [ 33 ]. Although a further experiment is required, Western blot results revealed that the developed 4 MAbs respond to the allergens Ara h 1 and 3 proteins widely recognized as TSSPs [ 34 ].…”

“…Heat treatment is the most representative method used in food industries to improve the shelf life and flavor of peanuts. It is well known that most proteins in food were turned to insoluble status and lost their antigenicity due to denaturation during the heat treatment, but some proteins have been reported to be able to maintain solubility and antigenicity [ 33 ].…”