Development of a β-Lactoglobulin Sensor Based on SPR for Milk Allergens Detection
A sensitive and label-free surface plasmon resonance (SPR) based sensor was developed in this work for the detection of milk allergens. β-lactoglobulin (BLG) protein was used as the biomarker for cow milk detection. This is to be used directly in final rinse samples of cleaning in-place (CIP) systems of food manufacturers. The affinity assay was optimised and characterised before a standard curve was performed in pure buffer conditions, giving a detection limit of 0.164 µg mL−1 as a direct binding assay. The detection limit can be further enhanced through the use of a sandwich assay and amplification with nanomaterials. However, this was not required here, as the detection limit achieved exceeded the required allergen detection levels of 2 µg mL−1 for β-lactoglobulin. The binding affinities of the polyclonal antibody for BLG, expressed by the dissociation constant (KD), were equal to 2.59 × 10−9 M. The developed SPR-based sensor offers several advantages in terms of label-free detection, real-time measurements, potential on-line system and superior sensitivity when compared to ELISA-based techniques. The method is novel for this application and could be applied to wider food allergen risk management decision(s) in food manufacturing.
Quantification of shellfish major allergen tropomyosin by SPR biosensor with gold patterned Biochips
Abstract Tropomyosin (TM), known as the major allergen in seafood, has been increasing concerns in industrialized countries. We proposed a simple, rapid and accurate analytical method using surface plasmon resonance (SPR) sensor with gold patterned biochips for detection and quantification of TM in shellfish. The limit of detection (LOD) was 1.0 μg mL−1 (S/N = 3), while the limit of quantitation (LOQ) was 2.5 μg mL−1 (S/N = 10). Moreover, the satisfactory recoveries (92.0–108.0%) was obtained for TM detection in food samples, showing the good accuracy of this method. Furthermore, the results also demonstrated that this developed method is suitable for the determination of TM from different shellfish species within 3 min. Therefore, the developed SPR sensor with gold patterned biochips will have positive significance on the rapid and accurate detection allergens in shellfish foodstuffs, related products and their production lines in the future.
Correlation between Fluoride Release, Microhardness and Solubility of High viscosity Glass Ionomers
This study was to measure and compare the amount of fluoride, microhardness and solubility of high viscosity glass ionomer and resin-reinforced glass ionomer during 84 days. Fuji IX GP EXTRA, Fuji IX GP, Fuji II LC and FiltekTM Z350XT stored in deionized water for 84 days to measure fluoride release, microhardness and solubility. As a result of measurement of fluoride release, all the glass ionomers showed the highest amount of fluoride release on day 1 and gradually decreased. Fuji IX GP EXTRA showed the highest amount of fluoride release and cumulative release. And Fuji IX GP and Fuji II LC showed no significant difference. Microhardness measurements showed that all experimental groups decreased 1 day after exposure to water. After 84 days, microhardness showed no significant difference between Fuji IX GP EXTRA and Fuji IX GP, and Fuji II LC was the lowest. In the solubility measurement, Fuji IX GP EXTRA, Fuji IX GP, and Fuji II LC increased rapidly to 21 days. After 21 days, there was no significant difference in the three groups. As a result, short term fluoride release affects solubility and microhardness, but long term fluoride release has no correlation. Through this study, the amount of fluoride, microhardness, and solubility of various glass ionomers were evaluated, and these properties could be applied clinically.