a 2022

ONE STEP CHEMICAL DERIVATIZATION OF SECRETED CELL METABOLITES FOR SENSITIVE MASS SPECTROMETRY DETECTION

PEČINKA, Lukáš, Lukáš MORÁŇ, Volodymyr POROKH, Martina MARCHETTI-DESCHMANN, Josef HAVEL et. al.

Basic information

Original name

ONE STEP CHEMICAL DERIVATIZATION OF SECRETED CELL METABOLITES FOR SENSITIVE MASS SPECTROMETRY DETECTION

Authors

PEČINKA, Lukáš, Lukáš MORÁŇ, Volodymyr POROKH, Martina MARCHETTI-DESCHMANN, Josef HAVEL, Aleš HAMPL and Petr VAŇHARA

Edition

XXI. Interdisciplinary meeting of young life scientists, 2022

Other information

Type of outcome

Konferenční abstrakt

Confidentiality degree

není předmětem státního či obchodního tajemství

References:

Organization unit

Faculty of Medicine
Změněno: 5/4/2023 13:08, Mgr. Tereza Miškechová

Abstract

V originále

Metabolomics studies for biomarker discovery consist of a complete set of metabolites in complex biological systems (databases contain ~ 115 000 metabolites). Metabolome is much smaller than proteome (1 800 000 different proteins) making it relatively simple for data analysis. On the other hand, metabolites are chemically diverse compounds that occur at vide range of concentrations (ten orders of magnitude) which makes metabolomics analysis challenging. Various chemical derivatization procedures have been extensively studied to enhance ionization efficacy, retention time (chromatography), and shift molecular mass to a higher value (mass spectrometry). Functional groups including aldehyde, primary amines, and carboxylic acid have been targeted using different derivatization reagents. Around 80% of a total number of described metabolites can be derivatized and detected by mass spectrometry using i.e. Girard´s reagent T – GT (aldehyde groups), Coniferyl aldehyde – CA (primary amines), and 2-picolylamine – 2-PA (carboxylic acids). Metabolomics methods are highly used to analyze large-scale phenotypic changes in the cell from intracellular (cell lysate) or extracellular (cell secretome) biological matrices. Cell cultivation process is accompanied by secretion of a broad range of metabolites into the cultivation media (i.e., autocrine and paracrine factors such as immune receptors, cytokines, and hormones) which are characteristic for individual cell types. Herein, the method for derivatization of various classes of small molecules has been optimized and compared to the detection sensitivity of non-derivatized molecules. The secreted metabolites profile of human embryonic stem cells (hESCs) has been analyzed through multiple derivatized procedures coupled with metabolomics with the aim of a better understanding of hESCs therapeutical effects.

Links

EF19_073/0016943, research and development project
Name: Interní grantová agentura Masarykovy univerzity
MUNI/A/1398/2021, interní kód MU
Name: Zdroje pro tkáňové inženýrství 12 (Acronym: TissueEng 12)
Investor: Masaryk University
MUNI/A/1421/2021, interní kód MU
Name: Efekt perioperační hemodynamické optimalizace na výskyt pooperačních komplikací u operací mozku. Monocentrická randomizovaná pilotní studie.
Investor: Masaryk University
MUNI/IGA/1127/2021, interní kód MU
Name: Molecular organization of the zygotic spindle apparatus in non-rodent embryos
Investor: Masaryk University
MUNI/IGA/1208/2021, interní kód MU
Name: Derivatization strategies revealing metabolic patterns in human pluripotent stem cells and progenitors by mass spectrometry (Acronym: Stem cells metabolites derivatization)
Investor: Masaryk University
MUNI/11/ACC/3/2022, interní kód MU
Name: Bioanalytical quality control of cGMP/ATMP-grade stem cells and progenitors
Investor: Masaryk University, Accelerate