Development of a β-Lactoglobulin Sensor Based on SPR for Milk Allergens Detection

被引:62
作者
Ashley, Jon [1 ,2 ]
D'Aurelio, Roberta [1 ]
Piekarska, Monika [1 ]
Temblay, Jeff [3 ]
Pleasants, Mike [3 ]
Trinh, Linda [4 ]
Rodgers, Thomas L. [4 ]
Tothill, Ibtisam E. [1 ]
机构
[1] Cranfield Univ, Adv Diagnost & Sensors Grp, Cranfield MK43 0AL, Beds, England
[2] Denmark Tech Univ, Dept Micro & Nanotechnol, DK-2800 Lyngby, Denmark
[3] Unilever Plc, Safety & Environm Assurance Ctr, Colworth Sci Pk, Bedford MK44 1LQ, England
[4] Univ Manchester, Sch Chem Engn & Analyt Sci, Oxford Rd, Manchester M13 9PL, Lancs, England
来源
BIOSENSORS-BASEL | 2018年 / 8卷 / 02期
基金
“创新英国”项目;
关键词
allergen; milk protein; beta-lactoglobulin (BLG); surface plasmon resonance (SPR); biosensor; QUARTZ-CRYSTAL MICROBALANCE; FOOD ALLERGENS; REAL-TIME; BIOSENSOR; QUANTIFICATION; CASEIN; QCM;
D O I
10.3390/bios8020032
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
A sensitive and label-free surface plasmon resonance (SPR) based sensor was developed in this work for the detection of milk allergens. beta-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 mu g 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 mu g mL(-1) for beta-lactoglobulin. The binding affinities of the polyclonal antibody for BLG, expressed by the dissociation constant (K-D), were equal to 2.59 x 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.
引用
收藏
页数:11
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