Enzyme-linked immunosorbent assay using fragment antigen-binding (Fab) antibody for determination of deoxymiroestrol, a potent phytoestrogen from Pueraria candollei

Deoxymiroestrol is the most potent phytoestrogen in chromenes group that has been found in Pueraria candollei, Thai name known as Kwao Krua Khao. Several studies reported estrogenic activity of P. candollei in order to using as hormone replacement therapy for postmenopausal women. Previously, specific determination of deoxymiroestrol content by enzyme-linked immunosorbent assay (ELISA) using polyclonal antibody (pAb) have been reported. However, production of pAb has limitation and variability from different batches. Therefore, in this study, we established quantitative method for determination of deoxymiroestrol using fragment antigen-binding (Fab) antibody-based immunoassay. The developed immunoassay has specificity to deoxymiroestrol with a calibration range of 15.6-1000 ng mL-1. Precision including intra-assay and inter-assay are 1.48-7.11 and 0.58-9.31%, respectively. Accuracy of the assay showed in recovery between 99.77-101.61% when spike deoxymiroestrol standard into the samples. The limit of detection (LOD) is 30.80 ng mL-1. Comparation antibody-based immunoassay for determination of deoxymiroestrol using Fab with pAb was represented consistency (R2 = 0.9807) when analysis roots bark of Pueraria candollei from difference areas. Therefore, this development assay can apply to determine deoxymiroestrol content in the plant samples.

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High performance enzyme-linked immunosorbent assay for determination of miroestrol, a potent phytoestrogen from Pueraria candollei

Analytica Chimica Acta ◽

10.1016/j.aca.2013.04.053 ◽

2013 ◽

Vol 785 ◽

pp. 104-110 ◽

Cited By ~ 15

Author(s):

Gorawit Yusakul ◽

Orapin Udomsin ◽

Thaweesak Juengwatanatrakul ◽

Hiroyuki Tanaka ◽

Chaiyo Chaichantipyuth ◽

...

Keyword(s):

Immunosorbent Assay ◽

High Performance ◽

Enzyme Linked Immunosorbent Assay ◽

Pueraria Candollei

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Determination of antigen binding specificities of Cryptococcus neoformans factor sera by enzyme-linked immunosorbent assay.

Infection and Immunity ◽

10.1128/iai.63.5.1810-1819.1995 ◽

1995 ◽

Vol 63 (5) ◽

pp. 1810-1819 ◽

Cited By ~ 22

Author(s):

T Belay ◽

R Cherniak

Keyword(s):

Cryptococcus Neoformans ◽

Immunosorbent Assay ◽

Enzyme Linked Immunosorbent Assay ◽

Antigen Binding

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EVALUATION OF A RECENTLY DEVELOPPED ENZYME LINKED IMMUNOSORBENT ASSAY FOR THE DETERMINATION OF ANTI-PHOSPHOLIPASE A2 RECEPTOR ANTIBODIES

10.26226/morressier.56e174d9d462b8028d88aca7 ◽

2016 ◽

Author(s):

Anne-Sophie Messiaen

Keyword(s):

Phospholipase A2 ◽

Immunosorbent Assay ◽

Enzyme Linked Immunosorbent Assay ◽

Phospholipase A2 Receptor ◽

Receptor Antibodies

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Biotin-avidin Mediated Competitive Enzyme-linked Immunosorbent Assay for Determination of Ketamine

CHINESE JOURNAL OF ANALYTICAL CHEMISTRY (CHINESE VERSION) ◽

10.3724/sp.j.1096.2010.00117 ◽

2010 ◽

Vol 38 (1) ◽

pp. 117-120

Author(s):

Wen-Di ZHANG ◽

Ping SU ◽

Yi YANG ◽

Zhen-Quan GUO

Keyword(s):

Immunosorbent Assay ◽

Enzyme Linked Immunosorbent Assay

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SENSITIVE AND SPECIFIC ENZYME-LINKED IMMUNOSORBENT ASSAY FOR UROKINASE-TYPE PLASMINOGEN ACTIVATOR IN HUMAN PLASMA

10.1055/s-0038-1644425 ◽

1987 ◽

Cited By ~ 1

Author(s):

J Grøndahl-HANSEN ◽

N Agerlin ◽

L S Nielsen ◽

K Danø

Keyword(s):

Plasminogen Activator ◽

Immunosorbent Assay ◽

Enzyme Linked Immunosorbent Assay ◽

Mean Values ◽

Amino Terminal ◽

Type Plasminogen Activator ◽

Healthy Donors ◽

Urokinase Type Plasminogen Activator ◽

The Mean

An enzyme-linked immunosorbent assay (ELISA) was developed for the measurement of human urokinase-type plasminogen activator (u-PA) in plasma and serum. Microtiter plates were coated with a monoclonal antibody and incubated with standard or sample. Bound u-PA was quantitated with polyclonal antibodies conjugated with biotin, followed by avidin-peroxidase. The assay was 10-fold as sensitive as other previously reported ELISAs, the detection limit being approximately 1 pg of u-PA in a volume of 100 μl with a linear dose-response up to 15 pg of u-PA. The assay detected active u-PA and its inactive proenzyme form equally well and the recovery of both forms was higher than 90% in plasma. A variety of structurally related proteins, including t-PA, were tested, but no reaction with proteins other than u-PA and its amino-terminal degradation product were observed. The intra-assay and inter-assay coefficients of variation for determination of u-PA in plasma were 7.6% and 8.4%, respectively. The assay was equally applicable to serum. The values obtained with plasma and serum were similar, and the results were not affected by small variations in the preparation of the samples. The ELISA was used to measure the concentration of u-PA in plasma from 34 healthy donors. The mean values for u-PA in plasma from healthy donors was 1.1 ng/ml ± 0.3 ng/ml (SD) (range 0.6 - 1.5 ng/ml). No significant differences were found between men and women and no correlation between u-PA concentration and age could be demonstrated.The mean u-PA concentration in plasma from healthy donors obtained in this study is substantially lower than that reported by others. This might be due to different methods of determination of the protein content of the standard preparations or to differences in the specificity of the assays.

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