Development of an enzyme-linked immunosorbent assay based on anti-puerarin monoclonal antibody and its applications

et al. 2015

Chin. J. Integr. Med.

Self Cite

mentioning

confidence: 99%

Effect of Puerarin on the Pharmacokinetics of Baicalin in Gegen Qinlian Decoction (葛根芩连汤) in Mice

Kong

et al. 2015

Chin. J. Integr. Med.

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“…The icELISA procedure to measure PUE was performed as described previously [15]. Briefly, the coating antigen PUE-BSA (0.1 µg/mL) was added to a microtitre plate and incubated.…”

Section: Icelisamentioning

confidence: 99%

Pharmacokinetics and Tissue Distribution Kinetics of Puerarin in Rats Using Indirect Competitive ELISA

Kong

Shi

et al. 2017

Preprint

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Puerarin (PUE) is a compound isolated from the roots of Pueraria lobata. We studied the pharmacokinetics and tissue distribution kinetics of PUE in Sprague-Dawley rats following intraperitoneal administration of three concentrations. Indirect competitive ELISA based on an anti-PUE monoclonal antibody was used to determine the concentration of PUE in the blood, heart, liver, spleen, lung, kidney, hippocampus, cerebral cortex, and striatum. The plasma and tissue distribution kinetic characteristics following a single injection of PUE (20, 40, and 80 mg/kg) were calculated using a non-compartment model. In the high-dose (80 mg/kg) and medium-dose (40 mg/kg) groups, the kinetic profile of PUE in blood and kidney samples showed two absorption peaks, while that of the other tissues showed only one peak. In the low-dose (20 mg/kg) group, there was only one peak, irrespective of the sample type. Pharmacokinetic parameters such as the area under the curve, Cmax, and Tmax varied according to the administered dose. AUC and Cmax values increased dose-dependently. PUE was widely distributed in areas of the brain such as the hippocampus, cerebral cortex, and striatum, providing a foundation for guiding the use of PUE in the treatment of cerebral ischaemic stroke and neurodegenerative diseases.

“…For this study, monoclonal antibodies against PF were purified by the caprylic acid-ammonium sulfate method [16] and protein G Sepharose 4 Fast Flow column (GE healthcare Bio-Sciences AB, Uppsala, Sweden) [12], and the antibody content was quantified using the Bradford Protein Assay Kit (Biotopped, Beijing, China).…”

Section: Purification Of Anti-pf Mabmentioning

confidence: 99%

“…In our ongoing study of the production of MAbs against naturally occurring bioactive compounds, we have produced MAbs for the detection of baicalin [10], geniposide [11], puerarin [12]. Using these MAbs, we have also developed indirect competitive enzyme-linked immunosorbent assays (icELISA) for the determination of these compounds in medicines and biological samples.…”

Section: Introductionmentioning

confidence: 99%

Development of Fluorescence-Linked Immunosorbent Assay for Paeoniflorin

Zhao

et al. 2015

J Fluoresc

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In this study, we developed a fluorescent immunoassay approach to detect paeoniflorin (PF) using a fluorescently labelled monoclonal antibody. The PF-specific antibody was purified by the caprylic acid-ammonium sulfate method and protein G Sepharose 4 Fast Flow column and then labelled with fluorescein isothiocyanate (FITC). The FITC-labelled monoclonal antibody was highly specific for PF, with less than 0.076 % cross-reactivity to seven structurally related compounds. The FITC-labelled monoclonal antibody was then used to develop an indirect competitive enzyme-linked immunosorbent assay (icELISA) and indirect competitive fluorescence-linked immunosorbent assay (icFLISA), respectively. FLISA is simple, rapid and sensitive, with a 500-fold lower limit of detection (LOD) compared with conventional ELISA. Finally, using a variety of standards, FLISA was validated. We observed a strong correlation between the results determined by either FLISA or conventional HPLC for the quantification of PF levels (R(2) = 0.9927). Collectively, this study shows that the icFLISA method can be successfully applied for the detection and quantification of PF in medicines and biological samples.