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Development of a β-Lactoglobulin Sensor Based on SPR for Milk Allergens Detection
被引:64
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.
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页数:11
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