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Development of a H2 O2 -sensitive quantum dots-based fluorescent sandwich ELISA for sensitive detection of bovine ß-lactoglobulin by monoclonal antibody.
He, Shengfa; Li, Xin; Gao, Jinyan; Tong, Ping; Chen, Hongbing.
Affiliation
  • He S; State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang, P.R. China.
  • Li X; School of Food Science & Technology, Nanchang University, Nanchang, P.R. China.
  • Gao J; Jiangxi-OAI Joint Research Institute, Nanchang University, Nanchang, P.R. China.
  • Tong P; State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang, P.R. China.
  • Chen H; School of Food Science & Technology, Nanchang University, Nanchang, P.R. China.
J Sci Food Agric ; 98(2): 519-526, 2018 Jan.
Article in En | MEDLINE | ID: mdl-28620918
ABSTRACT

BACKGROUND:

Bovine ß-lactoglobulin (BLG) is the major allergen in cows' milk, and the specific epitope plays a key role in food allergy. Developing a method specifically bind to the IgE epitope is necessary for testing BLG and its allergenic residues.

RESULTS:

The monoclonal antibody (1G9) specific to the IgE linear epitope for BLG was identified as high affinity and specificity. Based on 1G9, a sensitive fluorescent sandwich enzyme-linked immunosorbent assay (sELISA) was successfully developed using catalase-mediated fluorescence quenching of thiolated CdTe quantum dots in the presence of hydrogen peroxide as fluorescent signal output. The fluorescent sELISA showed high sensitivity and specificity, the limit of detection was 0.49 ng mL-1 , which was 16-fold lower than horseradish peroxidase (HRP)-based sELISA. The linear range for BLG detection were 125-4000 ng mL-1 (r = 0.9939) and 0.48-62.5 ng mL-1 (r = 0.9919). The recoveries and coefficients of variation were 94.25-109.83% and 4.38-20.29%, respectively. Allergenic residues were also detected in hydrolysed infant formulas. The results of fluorescent sELISA showed good performance as HRP-based sELISA and commercial sELISA kit.

CONCLUSION:

This proposed fluorescent sELISA could be employed to detect BLG and its allergenic residues in food with highly sensitivity, reliability, and recovery. © 2017 Society of Chemical Industry.
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Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: Enzyme-Linked Immunosorbent Assay / Hydrogen Peroxide / Lactoglobulins Type of study: Diagnostic_studies / Evaluation_studies Limits: Animals / Humans Language: En Journal: J Sci Food Agric Year: 2018 Document type: Article

Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: Enzyme-Linked Immunosorbent Assay / Hydrogen Peroxide / Lactoglobulins Type of study: Diagnostic_studies / Evaluation_studies Limits: Animals / Humans Language: En Journal: J Sci Food Agric Year: 2018 Document type: Article