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Immunoassay for determination of trilobolide
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SYSNO ASEP 0474379 Druh ASEP J - Článek v odborném periodiku Zařazení RIV J - Článek v odborném periodiku Poddruh J Článek ve WOS Název Immunoassay for determination of trilobolide Tvůrce(i) 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)Zdroj.dok. Steroids. - : Elsevier - ISSN 0039-128X
Roč. 117, Jan (2017), s. 105-111Poč.str. 7 s. Akce Conference on Isoprenoids /23./ Datum konání 04.09.2016 - 07.09.2016 Místo konání Minsk Země BY - Bělorusko Typ akce WRD Jazyk dok. eng - angličtina Země vyd. US - Spojené státy americké Klíč. slova trilobolide ; avidin-biotin ; ELISA ; Laser trilobum ; synthesis ; immunoassay Vědní obor RIV CE - Biochemie Obor OECD Biochemical research methods Vědní obor RIV – spolupráce Ústav experimentální medicíny - Farmakologie a lékárnická chemie Institucionální podpora UOCHB-X - RVO:61388963 ; UEM-P - RVO:68378041 UT WOS 000392566800015 EID SCOPUS 84995488385 DOI 10.1016/j.steroids.2016.08.019 Anotace 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. Pracoviště Ústav organické chemie a biochemie Kontakt asep@uochb.cas.cz ; Kateřina Šperková, Tel.: 232 002 584 ; Viktorie Chládková, Tel.: 232 002 434 Rok sběru 2018
Počet záznamů: 1