TY - JOUR
T1 - Open sandwich fluorescence-linked immunosorbent assay for detection of soy isoflavone glycosides
AU - Sakamoto, Seiichi
AU - Uchiyama, Hiroki
AU - Yusakul, Gorawit
AU - Kyokong, Napat
AU - Pongkitwitoon, Benyakan
AU - Putalun, Waraporn
AU - Tanaka, Hiroyuki
AU - Morimoto, Satoshi
N1 - Funding Information:
This work was supported in part by a grant from Fuji Foundation for Protein Research. We appreciate the technical support from the Research Support Center, Graduate School of Medical Sciences, Kyushu University.
Publisher Copyright:
© 2021 Elsevier Ltd
PY - 2021/11/1
Y1 - 2021/11/1
N2 - To detect major soy isoflavone glycosides, namely daidzin (DZ) and genistin (GEN), novel open sandwich fluorescence-linked immunosorbent assay (os-FLISA) was developed by taking advantage of enhanced interactions between variable regions of heavy (VH) and light chain (VL) domains in the presence of an antigen. The VH and VL genes were expressed in Escherichia coli as a chimera protein with green fluorescence protein (AcGFP1) and maltose-binding protein (MBP), respectively. Comprehensive characterization of os-FLISA displayed nearly the same specificity as parental DZ- and GEN-specific monoclonal antibody, demonstrating the potential of the developed assay for detection of both DZ and GEN. Their detectable range in this system exhibited at 0.1–12.5 μg mL−1. Subsequent validation analysis revealed that os-FLISA was reliable and accurate system for detection of total soy isoflavone glycosides. Notably, this is the first FLISA based on an open sandwich system, which can be employed for the detection of small molecules.
AB - To detect major soy isoflavone glycosides, namely daidzin (DZ) and genistin (GEN), novel open sandwich fluorescence-linked immunosorbent assay (os-FLISA) was developed by taking advantage of enhanced interactions between variable regions of heavy (VH) and light chain (VL) domains in the presence of an antigen. The VH and VL genes were expressed in Escherichia coli as a chimera protein with green fluorescence protein (AcGFP1) and maltose-binding protein (MBP), respectively. Comprehensive characterization of os-FLISA displayed nearly the same specificity as parental DZ- and GEN-specific monoclonal antibody, demonstrating the potential of the developed assay for detection of both DZ and GEN. Their detectable range in this system exhibited at 0.1–12.5 μg mL−1. Subsequent validation analysis revealed that os-FLISA was reliable and accurate system for detection of total soy isoflavone glycosides. Notably, this is the first FLISA based on an open sandwich system, which can be employed for the detection of small molecules.
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U2 - 10.1016/j.foodchem.2021.129829
DO - 10.1016/j.foodchem.2021.129829
M3 - Article
C2 - 34087571
AN - SCOPUS:85107679668
SN - 0308-8146
VL - 361
JO - Food Chemistry
JF - Food Chemistry
M1 - 129829
ER -