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

Mei Shu, Yang Xu, Xing Liu, Yanping Li, Qinghua He, Zhui Tu, Jinheng Fu, Shirley J. Gee, Bruce D. Hammock

Research output: Contribution to journalArticlepeer-review

37 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)53-59
Number of pages7
JournalAnalytica Chimica Acta
Volume924
DOIs
StatePublished - Jun 14 2016

Keywords

  • Anti-idiotypic antibody
  • Nanobody
  • One-step immunoassay

ASJC Scopus subject areas

  • Analytical Chemistry
  • Environmental Chemistry
  • Biochemistry
  • Spectroscopy

Fingerprint Dive into the research topics of 'Anti-idiotypic nanobody-alkaline phosphatase fusion proteins: Development of a one-step competitive enzyme immunoassay for fumonisin B<sub>1</sub> detection in cereal'. Together they form a unique fingerprint.

Cite this