Number of the records: 1
Immunoassay for determination of trilobolide
- 1.
SYSNO ASEP 0474379 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title Immunoassay for determination of trilobolide Author(s) Huml, L. (CZ)
Jurášek, M. (CZ)
Mikšátková, P. (CZ)
Zimmermann, T. (CZ)
Tomanová, P. (CZ)
Buděšínský, Miloš (UOCHB-X) RID, ORCID
Rottnerová, Z. (CZ)
Šimková, M. (CZ)
Harmatha, Juraj (UOCHB-X) RID, ORCID
Kmoníčková, Eva (UEM-P) RID
Lapčík, O. (CZ)
Drašar, P. B. (CZ)Source Title Steroids. - : Elsevier - ISSN 0039-128X
Roč. 117, Jan (2017), s. 105-111Number of pages 7 s. Action Conference on Isoprenoids /23./ Event date 04.09.2016 - 07.09.2016 VEvent location Minsk Country BY - Belarus Event type WRD Language eng - English Country US - United States Keywords trilobolide ; avidin-biotin ; ELISA ; Laser trilobum ; synthesis ; immunoassay Subject RIV CE - Biochemistry OECD category Biochemical research methods Subject RIV - cooperation Institute of Experimental Medicine - Pharmacology ; Medidal Chemistry Institutional support UOCHB-X - RVO:61388963 ; UEM-P - RVO:68378041 UT WOS 000392566800015 EID SCOPUS 84995488385 DOI 10.1016/j.steroids.2016.08.019 Annotation Trilobolide (Tb) is a pharmacologically interesting sesquiterpene lactone isolated from Laser trilobum (L.) Borkh. Structural relation to a sarco/endoplasmic reticulum Ca2+-ATPase inhibitor thapsigargin bring promising prospects for Tb to be used in the development of new anti-cancer drugs. As long as there are still unanswered questions regarding its investigation, a need for novel analytical tools emerge. Since immunoassays serve as one of powerful tools within the investigation of natural products, the development of indirect competitive enzyme-linked immunosorbent assay (ELISA) utilizing coating based on avidin-biotin technology is described. In our set-up of ELISA, newly synthesized biotinylated Tb served as immobilized competitor. Tb-carboxymethyloxime-bovine serum albumin (BSA) and Tb-succinoyl-BSA conjugates were used separately for immunization of rabbits. Two sets of polyclonal antibodies (RAbs) were obtained. Antibodies against Tb-succinoyl-BSA conjugate (RAb No. 206) were chosen as the best. Under optimized conditions, limit of detection and 50% intercept of our ELISA were 849 pg/mL and 8.89 ng/mL, respectively. The cross-reactivity (CR) was tested on 10 structurally related compounds and CR did not exceed 6.1%. The reproducibility of the system is expressed as intra- and inter-assay coefficients of variation (9.7% and 11.4%, respectively). Based on conducted experiments, we proposed the use of ELISA for quantification of Tb in complex biological matrices such as plant extracts. A method was applied to analyze three extracts obtained from different parts of L trilobum. Data obtained were compared to those acquired by UHPLC-MS/MS. The concordance between the methods (103-87%) showed the ability of ELISA to quantify Tb. Workplace Institute of Organic Chemistry and Biochemistry Contact asep@uochb.cas.cz ; Kateřina Šperková, Tel.: 232 002 584 ; Viktorie Chládková, Tel.: 232 002 434 Year of Publishing 2018
Number of the records: 1