Imaging surface plasmon resonance for multiplex microassay sensing of mycotoxins

Abstract

A prototype imaging surface plasmon resonance-based multiplex microimmunoassay for mycotoxins is described. A microarray of mycotoxin–protein conjugates was fabricated using a continuous flow microspotter device. A competitive inhibition immunoassay format was developed for the simultaneous detection of deoxynivalenol (DON) and zearalenone (ZEN), using a single sensor chip. Initial in-house validation showed limits of detection of 21 and 17 ng/mL for DON and 16 and 10 ng/mL for ZEN in extracts, which corresponds to 84 and 68 μg/kg for DON and 64 and 40 μg/kg for ZEN in maize and wheat samples, respectively. Finally, the results were critically compared with data obtained from liquid chromatography-mass spectrometry confirmatory analysis method and found to be in good agreement. The described multiplex immunoassay for the rapid screening of several mycotoxins meets European Union regulatory limits and represents a robust platform for mycotoxin analysis in food and feed samples.

Colour-encoded paramagnetic microbead-based direct inhibition triplex flow cytometric immunoassay for ochratoxin A, fumonisins and zearalenone in cereals and cereal-based feed

“…Apparently, the epitopes of the conjugated ZEN, α-ZEL and β-ZEL molecules used in our research are blocked for antibody recognition. These results are comparable to Dorokhin et al [30] who used the same antibody in a label-free iSPR approach, except the result for β-ZEL, which differs substantially (10-fold). This might be due to the entirely different technique and the indirect assay approach used in that work.…”

“…A new approach is the multiplex detection of mycotoxins using the label-free imaging SPR (iSPR) technique. Using an IBIS iSPR, capable of reading out a spotted microarray sensor chip, Dorokhin et al [30] developed a method for the simultaneous detection of ZEN and DON. The used iSPR technology facilitates higher multiplexing capacity as was shown for allergens [31].…”

Affinity maturation of single-chain variable fragment specific for aflatoxin B1 using yeast surface display

“…However, such chromatographic methods are inappropriate as an on-site detection system especially in developing countries due to time-consuming and cost-intensiveness. Therefore, the requirement for fast, portable and convenient detection tools of AFB 1 has led to the development of immunological methods such as enzyme-linked immunosorbent assay (ELISA), immuno-chromatographic strip or protein chip (Cho et al, 2005;Dorokhin, Haasnoot, Franssen, Zuilhof, & Nielen, 2011;Park, Kim, Kim, & Ko, 2014). So far monoclonal as well as polyclonal antibodies specific for AFB 1 have been used for various immunological detection modules and there are many commercialized kits (Groopman, Trudel, Donahue, Marshak-Rothstein, & Wogan, 1984;Haugen et al, 1981;Liu, Hsu, Lu, & Yu, 2013;Martin et al, 1984) which absolutely depend on their specificity and sensitivity.…”

Application of Modern Biosensors Methods in Ecotoxicological Monitoring of Some Toxins of Natural (Micotoxins) and Antropogenic (Pesticides) Origin. Part 1. Micotoxins.

“…Для одночасного виявлення деоксинівале-нолу та зеараленону Dorokhin et al [120] роз-вивали конкурентний метод мультикомплек-сного мікроіммунологічного обстеження ви-конуючи ППР у форматі відображення (imaging) (iSPR). Мікровипробування, проведені на карбоксильованій поверхні чіпа сенсора (Biacor 3000) та вимірювання, забезпечені IBIS iSPR, показали межу виявлення 21 і 17 нг/мл для ДОН і 16 і 10 нг/мл для ЗЕН, в екстрактах, які відповідають 84 і 68 мкг/кг для ДОН і 64 і 40 мкг/кг для ЗЕН в кукурудзі і зразках пшени-ці, відповідно.…”