Development of Fluorescence-Linked Immunosorbent Assay for Paeoniflorin.
J Fluoresc
; 25(4): 885-90, 2015 Jul.
Article
in En
| MEDLINE
| ID: mdl-25895576
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.
Full text:
1
Database:
MEDLINE
Traditional Medicines:
Medicinas_tradicionales_de_asia
/
Medicina_china
Main subject:
Enzyme-Linked Immunosorbent Assay
/
Drugs, Chinese Herbal
/
Monoterpenes
/
Glucosides
/
Immunosorbents
/
Antibodies, Monoclonal
Language:
En
Journal:
J Fluoresc
Year:
2015
Type:
Article
Affiliation country:
China