Preparation of Specific Nanobodies and Their Application in the Rapid Detection of Nodularin-R in Water Samples

Abstract

Nanobodies have several advantages, including great stability, sensibility, and ease of production; therefore, they have become important tools in immunoassays for chemical contaminants. In this manuscript, nanobodies for the detection of the toxin Nodularin-r (NOD-R), a secondary metabolite of cyanobacteria that could cause a safety risk for drinks and food for its strong hepatotoxicity, were for the first time selected from an immunized Bactrian camel VHH phage display library. Then, a sensitive indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) for NOD-R, based on the nanobody N56 with great thermostability and organic solvent tolerance, was established under optimized conditions. The results showed that the limit of detection for NOD-R was 0.67 µg/L, and the average spike recovery rate was between 84.0 and 118.3%. Moreover, the ic-ELISA method was validated with spiked water sample and confirmed by UPLC–MS/MS, which indicated that the ic-ELISA established in this work is a reproducible detection assay for nodularin residues in water samples.

Preparation of Novel Nanomaterial and Its Application in Food Industry

“…The results of the two methods are basically the same. In addition, the nanobody, as an important tool in immunoassay for chemical contaminants, was developed and its efficiency was examined by detecting a secondary metabolite of cyanobacteria, namely nodularin (NOD-R) [11]. The ic-ELISA method based on the nanobody N56 was validated with spiked water sample and confirmed by UPLC-MS/MS, which indicated that the ic-ELISA established in this work is a reproducible detection assay for nodularin residues in water samples.…”

Food-Borne Biotoxin Neutralization in Vivo by Nanobodies: Current Status and Prospects

“…NOD-R is a characteristic cyclic peptide composed of five amino acids. The first screening of Nbs against this cyanobacterial secondary metabolite toxin was conducted by Yang et al 104 However, these studies have also not yet been correlated with in vivo neutralization. Apart from the toxins mentioned above, there are also some common phytotoxins such as phytohemagglutinin, Capsule Lotus Root toxin, Solanine, and coniine.…”

Production of nanobodies in Andean camelids and their most common applications: A general review in the medical field.

“…Nanobodies used in water and soil allow the quick and accurate identification and detection of toxins and contaminants to the nanogram levels so, becoming a handy tool for these purposes [130][131][132] .…”