TY - JOUR
T1 - An indirect competitive enzyme-linked immunosorbent assay toward the standardization of Pueraria candollei based on its unique isoflavonoid, kwakhurin
AU - Yusakul, Gorawit
AU - Togita, Rui
AU - Minami, Kei
AU - Chanpokapaiboon, Kaskamol
AU - Juengwatanatrakul, Thaweesak
AU - Putalun, Waraporn
AU - Tanaka, Hiroyuki
AU - Sakamoto, Seiichi
AU - Morimoto, Satoshi
N1 - Funding Information:
The authors acknowledge the financial support from the Thailand Research Fund and Office of the Higher Education Commission (grant number MRG6180044 ). The authors thank Dr. Chaiyo Chaichantipyuth, Faculty of Pharmaceutical Sciences, Chulalongkorn University , Bangkok, Thailand, for providing authentic Kwa.
Funding Information:
The authors acknowledge the financial support from the Thailand Research Fund and Office of the Higher Education Commission (grant number MRG6180044). The authors thank Dr. Chaiyo Chaichantipyuth, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok, Thailand, for providing authentic Kwa.
Publisher Copyright:
© 2018 Elsevier B.V.
PY - 2019/3
Y1 - 2019/3
N2 - Kwakhurin (Kwa) is a plant secondary metabolite solely present in Pueraria candollei var. mirifica (P. candollei), which has long been used as a Thai traditional herb for estrogen replacement therapy. Recently, health hazards have arisen in Japan regarding P. candollei-derived products containing potent estrogenic compounds. Therefore, the development of standardization methods for P. candollei materials is an urgent problem requiring resolution. The enzyme-linked immunosorbent assay (ELISA) is an effective analytical technique because it enables the development of sensitive and specific assays of the target compound through antigen−antibody reaction. Here, we produced a monoclonal antibody against Kwa (MAb 11F) by immunizing Kwa-bovine serum albumin (BSA) conjugates prepared using an N,N′-carbonyldiimidazole (CDI) mediated method. Stability and cross-reactivity tests of MAb 11F revealed that the MAb 11F is stable for at least 4 months at 4 °C and is highly specific to Kwa. The detectable concentration range of an indirect competitive ELISA (icELISA) using MAb 11F exhibited values of 1.53–48.8 ng/mL with the limit of detection (LOD) of 1.13 ng/mL. Validation analyses revealed that the developed icELISA is precise, accurate, and reliable enough to be applied to P. candollei-derived samples and products for their standardization.
AB - Kwakhurin (Kwa) is a plant secondary metabolite solely present in Pueraria candollei var. mirifica (P. candollei), which has long been used as a Thai traditional herb for estrogen replacement therapy. Recently, health hazards have arisen in Japan regarding P. candollei-derived products containing potent estrogenic compounds. Therefore, the development of standardization methods for P. candollei materials is an urgent problem requiring resolution. The enzyme-linked immunosorbent assay (ELISA) is an effective analytical technique because it enables the development of sensitive and specific assays of the target compound through antigen−antibody reaction. Here, we produced a monoclonal antibody against Kwa (MAb 11F) by immunizing Kwa-bovine serum albumin (BSA) conjugates prepared using an N,N′-carbonyldiimidazole (CDI) mediated method. Stability and cross-reactivity tests of MAb 11F revealed that the MAb 11F is stable for at least 4 months at 4 °C and is highly specific to Kwa. The detectable concentration range of an indirect competitive ELISA (icELISA) using MAb 11F exhibited values of 1.53–48.8 ng/mL with the limit of detection (LOD) of 1.13 ng/mL. Validation analyses revealed that the developed icELISA is precise, accurate, and reliable enough to be applied to P. candollei-derived samples and products for their standardization.
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U2 - 10.1016/j.fitote.2018.12.010
DO - 10.1016/j.fitote.2018.12.010
M3 - Article
C2 - 30572086
AN - SCOPUS:85058674994
SN - 0367-326X
VL - 133
SP - 23
EP - 28
JO - Fitoterapia
JF - Fitoterapia
ER -