Development of Fluorescence-Linked Immunosorbent Assay for Paeoniflorin

Abstract

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.

Monoclonal Antibodies and Immunoassay for Medical Plant-Derived Natural Products: A Review

“…The MAbs against paeoniflorin was labelled with FITC and then used to develop an indirect competitive FLISA (icFLISA), which can be successfully applied for the detection and quantification of paeoniflorin in medicines and biological samples [ 68 ].…”

Development of a Competitive Time‐Resolved Fluoroimmunoassay for Paclitaxel

“…We previously tested the effectiveness of immunoassay in quality control of several natural medicines (Fujii et al ., ; Tung et al ., ; Oiso et al ., ; Li et al ., ; Zhao et al ., ); in fact, the earliest attempt of using immunoassay for Tax detection was reported over 20 years ago (Grothaus et al ., , ; Jaziri et al ., ; Leu et al ., , ; Svojanovsky et al ., ). Recently, we successfully prepared an anti‐Tax monocolonal antibody (MAb) 3A3, based on which we established an indirect competitive enzyme‐linked immunosorbent assay (icELISA) for Tax determination with a wide detection range of 9.8 to 312.5 ng/mL, showing obvious advantages over the previously reported methods (Chao et al ., ).…”

Pharmacokinetic analysis of orally administered puerarin in human saliva using an indirect competition ELISA

“…28 Furthermore, the IAC we developed based on the anti-ginsenoside Rh1 MAb was demonstrated as a potential method for the separation of epimers. We also succeeded in developing a uorescence-linked immunosorbent assay for paeoniorin, 31 which is more sensitive and time-saving.…”