Development and Characterization of Monoclonal Antibodies for the Mycotoxin Citreoviridin.

Chris M Maragos,Naoki Kobayashi,Yoshiko Sugita-Konishi,Yosuke Uchiyama,Fumichika Kominato

doi:10.3390/toxins11110630

Abstract

Citreoviridin (CTV) in an inhibitor of mitochondrial ATPase that has been isolated from molded yellow rice and linked to the human disease Shoshin-kakke (acute cardiac beriberi). The disease results from a deficiency of thiamine, however, purified CTV can reproduce the symptoms in experimental animals. The link between CTV and Shoshin-kakke has been difficult to resolve, in part because cases of the disease are rare. In addition to rice, CTV has been found in maize, pecan nuts, and wheat products. A method to screen for CTV and its geometric isomer, iso-CTV, in commodities was developed, based upon the isolation of two novel monoclonal antibodies (mAb). In an antigen-immobilized competitive enzyme-linked immunosorbent assay format (CI-ELISA), the observed IC50s for CTV were 11 ng/mL and 18 ng/mL (mAbs 2-2 and 2-4, respectively). The assays were relatively tolerant to methanol and acetonitrile, which allowed their application to the detection of CTV in spiked polished white rice. For quantification, a standard mixture of CTV and iso-CTV was used, along with matrix matched calibration. The dynamic range of the ELISA using mAb 2-4 was equivalent to 0.23 to 2.22 mg/kg in rice. Recoveries over the range of 0.36 to 7.23 mg/kg averaged 97 ± 10%. The results suggest that the mAb 2-4-based immunoassay can be applied to the screening of white rice for CTV. Both mAbs were also observed to significantly enhance the fluorescence of the toxin.

Highlights

In Japan in the years before World War I and continuing through the 1920s, there were human illnesses associated with consumption of moldy, yellow, rice
The results suggest that the monoclonal antibodies (mAb) 2-4-based immunoassay can be applied to the screening of white rice for CTV
Antibodies produced by11, these cell lines were evaluated in a CI-ELISA format for their sensitivity towards free x FORtwo

Summary

Introduction

In Japan in the years before World War I and continuing through the 1920s, there were human illnesses associated with consumption of moldy, yellow, rice. The illness, classified at the time as Shoshin-kakke (acute cardiac beriberi) decreased in incidence significantly around 1910, a fact attributed to increased inspection of rice by Japanese authorities [1]. The disease was related to beriberi, known to be caused by thiamine deficiency. By 1930 Shoshin-kakke had almost completely disappeared from Japan [1]. Subsequent investigations in Japan led to the differentiation of the broad category of yellow-colored rice into five groups, of which four are caused by Penicillium spoilage and one by Eurotium amstelodami [2]. The four caused by Penicillia are each associated with a different species

Methods

Results

Conclusion