Anti-idiotypic nanobody-alkaline phosphatase fusion proteins: Development of a one-step competitive enzyme immunoassay for fumonisin B1 detection in cereal.

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Abstract

A rapid and sensitive one-step competitive enzyme immunoassay for the detection of FB1 was developed. The anti-idiotypic nanobody-alkaline phosphatase (Ab2β-Nb-AP) was validated by the AP enzyme activity and the properties of bounding to anti-FB1-mAb (3F11) through colorimetric and chemiluminescence analyses. The 50% inhibitory concentration and the detection limit (LOD) of colorimetric enzyme-linked immunosorbent assay (ELISA) for FB1 were 2.69 and 0.35 ng mL(-1), respectively, with a linear range of 0.93-7.73 ng mL(-1). The LOD of the chemiluminescence ELISA (CLIA) was 0.12 ng mL(-1), and the IC50 was 0.89 ± 0.09 ng mL(-1) with a linear range of 0.29-2.68 ng mL(-1). Compared with LC-MS/MS, the results of this assay indicated the reliability of the Ab2β-Nb-AP fusion protein based one-step competitive immunoassay for monitoring FB1 contamination in cereals. The Ab2β-Nb-AP fusion proteins have the potential to replace chemically-coupled probes in competitive enzyme immunoassay systems.

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Author and article information

Journal

Journal ID (iso-abbrev): Anal. Chim. Acta

Title: Analytica chimica acta

Publisher: Elsevier BV

ISSN (Electronic): 1873-4324

ISSN (Print): 0003-2670

Publication date (Electronic): Jun 14 2016

Volume: 924

Affiliations

[1 ] State Key Laboratory of Food Science and Technology, Nanchang University, No. 235 Nanjing East Road, Nanchang 330047, People's Republic of China; Jiangxi-OAI Joint Research Institute, Nanchang University, No. 235 Nanjing East Road, Nanchang 330047, People's Republic of China.

[2 ] State Key Laboratory of Food Science and Technology, Nanchang University, No. 235 Nanjing East Road, Nanchang 330047, People's Republic of China; Jiangxi-OAI Joint Research Institute, Nanchang University, No. 235 Nanjing East Road, Nanchang 330047, People's Republic of China. Electronic address: xuyang@ncu.edu.cn.

[3 ] State Key Laboratory of Food Science and Technology, Nanchang University, No. 235 Nanjing East Road, Nanchang 330047, People's Republic of China; College of Food Science and Technology, Hainan University, No. 58 Renmin Avenue, Haikou 570228, China.

[4 ] Jiangxi-OAI Joint Research Institute, Nanchang University, No. 235 Nanjing East Road, Nanchang 330047, People's Republic of China.

[5 ] State Key Laboratory of Food Science and Technology, Nanchang University, No. 235 Nanjing East Road, Nanchang 330047, People's Republic of China.

[6 ] Department of Entomology and UCD Comprehensive Cancer Center, University of California, Davis, CA 95616, USA.

Article

Publisher Item ID: S0003-2670(16)30403-2

DOI: 10.1016/j.aca.2016.03.053

PubMed ID: 27181644

SO-VID: 0c619f74-5d47-4d4e-a22b-3bf06dac9b13

History

Keywords: Anti-idiotypic antibody,Nanobody,One-step immunoassay

Data availability:

Keywords: Anti-idiotypic antibody, Nanobody, One-step immunoassay

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