A sensitive and specific indirect competitive enzyme-linked immunosorbent assay for the detection of icariin

Abstract

In the present study, a novel monoclonal antibody (MAb) specific for icariin (ICA) was prepared and characterized. A hybridoma‑secreting MAb against icariin was produced by fusing splenocytes immunized with an ICA‑bovine serum albumin conjugate with a hypoxanthine‑aminopterin‑thymidine‑sensitive mouse myeloma SP2/0 cell line. The antibody showed high specificity for ICA with almost no cross‑reactivity against the majority of structurally‑related chemicals. Subsequently, an indirect competitive enzyme‑linked immunosorbent assay (ELISA) for ICA was established and characterized. In this assay, an effective measuring range of 10‑1,000 ng/ml of ICA (R2=0.9828) was detected. Intra‑ and inter‑assay repeatability and precision were achieved with a relative standard deviation (RSD) of <10%. A mean recovery of 95‑115% was obtained, with an RSD of <10%. In addition, the levels of ICA in traditional Chinese herbal prescriptions were determined, and correlation between the ELISA and high‑performance liquid chromatography analyses of total ICA was obtained. These results demonstrated that a reliable ELISA method had been successfully developed to determine ICA in traditional Chinese herbs and may contribute to further clinical investigations.

The Effects of Sweet Foods on the Pharmacokinetics of Glycyrrhizic Acid by icELISA

“…In contrast, enzyme linked immunosorbent assay (ELISA) is established by using a monoclonal antibody, its advantages include: specificity, sensitivity, repeatability, simple pretreatment of sample, fast and simple operation, economic and no-polluting, etc. Therefore, it has received more and more attention [ 14 , 15 , 16 , 17 , 18 ]. It is capable of analyzing large number of samples simultaneously, especially in cases where there is a very low content of the active ingredient.…”