| Texto completo | |
| Autor(es): |
Seidl, Claudia
;
de Lima, Juliana Maria
;
Leme, Gabriel Mazzi
;
Pires, Ananda Ferreira
;
Stoll, Dwight R.
;
Cardoso, Carmen Lucia
Número total de Autores: 6
|
| Tipo de documento: | Artigo Científico |
| Fonte: | FRONTIERS IN MOLECULAR BIOSCIENCES; v. 9, p. 14-pg., 2022-03-16. |
| Resumo | |
The continuous interest in discovering new bioactive molecules derived from natural products (NP) has stimulated the development of improved screening assays to help overcome challenges in NP-based drug discovery. Here, we describe a unique platform for the online screening of acetylcholinesterase inhibitors without the need for pre-treating the sample. In the current study, we have demonstrated the ability to combine reversed-phase separation with a capillary immobilized enzyme reactor (cIMER) in two-dimensional liquid chromatography system coupled with mass spectrometry detection. We systematically investigated the effects of method parameters that are of practical significance and are known to affect the enzyme assay and interfere in the analysis such as: bioreactor dimensions, loop sizes, amount of immobilized enzyme, second dimension flow rates, reaction time, substrate concentration, presence of organic modifier, limit of detection and signal suppression. The performance of this new platform was evaluated using a mixture containing three known AChE inhibitors (tacrine, galanthamine and donepezil) and an ethanolic extract obtained from the dry bulbs of Hippeastrum calyptratum (Amaryllidaceae) was investigated to provide a proof of concept of the applicability of the platform for the analysis of complex mixtures such as those derived from NPs. (AU) | |
| Processo FAPESP: | 14/50249-8 - Green chemistry: sustainable synthetic methods employing benign solvents, safer reagents, and bio-renewable feedstock |
| Beneficiário: | Arlene Gonçalves Corrêa |
| Modalidade de apoio: | Auxílio à Pesquisa - Programa Centros de Pesquisa em Engenharia |
| Processo FAPESP: | 14/11640-3 - Métodos LC-MS/2D para triagem de ligantes para a avaliação de atividade inibitória de enzimas e identificação dos compostos ativos em extratos brutos |
| Beneficiário: | Cláudia Seidl |
| Modalidade de apoio: | Bolsas no Brasil - Pós-Doutorado |
| Processo FAPESP: | 14/06907-0 - Imobilização da enzima nucleosídeo difosfato quinase (NDK) de Leishmania major: do estudo de condições à aplicação na triagem de ligantes |
| Beneficiário: | Juliana Maria de Lima |
| Modalidade de apoio: | Bolsas no Brasil - Doutorado Direto |
| Processo FAPESP: | 19/05363-0 - Triagem de ligantes enzimáticos: uma abordagem on-flow via LC-MS/MS |
| Beneficiário: | Carmen Lúcia Cardoso |
| Modalidade de apoio: | Auxílio à Pesquisa - Regular |
| Processo FAPESP: | 16/02941-5 - Desenvolvimento e otimização de metodologias bidimensionais de cromatografia líquida acopladas a bioensaios para a descoberta de inibidores enzimáticos em extratos de produtos naturais |
| Beneficiário: | Cláudia Seidl |
| Modalidade de apoio: | Bolsas no Exterior - Estágio de Pesquisa - Pós-Doutorado |