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001-es BibID:BIBFORM092956
Első szerző:Fonslow, Bryan
Cím:Multilevel Characterization of Antibody-Ligand Conjugates by CESI-MS / Bryan Fonslow, Gábor Jarvas, Marton Szigeti, András Guttman
Dátum:2020
ISSN:1566-5240
Megjegyzések:Aim: To demonstrate the capabilities of our new capillary electrophoresis mass spectrometry method, which facilitates highly accurate relative quantitation of modification site occupancy of antibody-ligand (e.g., antibody-drug) conjugates. Background: Antibody-drug conjugates play important roles in medical discovery for imaging and therapeutic intervention. The localization and stoichiometry of the conjugation can affect the orientation, selectivity, specificity, and strength of molecular interactions, influencing biochemical function. Objective: To demonstrate the option to analyze the localization and stoichiometry of antibody-ligand conjugates by using essentially the same method at all levels including ligand infusion, peptide mapping, as well as reduced and intact protein analysis. Materials and Methods: Capillary electrophoresis coupled with electrospray ionization mass spectrometry was used to analyze the antibody-ligand conjugates. Results: We identified three prevalent ligand conjugation sites with estimated stoichiometries of 73, 14, and 6% and an average ligand-antibody ratio of 1.37, illustrating the capabilities of CE-ESI-MS for rapid and efficient characterization of antibody-drug conjugates. Conclusion: The developed multilevel analytical method offers a comprehensive way to determine the localization and stoichiometry of antibody-drug conjugates for molecular medicinal applications. In addition, a significant advantage of the reported approach is the small, hydrophilic, unmodified peptides well separated from the neutrals, which is not common with other liquid phase separation methods such as LC.
Tárgyszavak:Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban
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Megjelenés:Current Molecular Medicine. - 20 : 10 (2020), p. 789-797. -
További szerzők:Járvás Gábor (1982-) (vegyészmérnök) Szigeti Márton (1986-) (környezetmérnök) Guttman András (1954-) (vegyészmérnök)
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001-es BibID:BIBFORM092960
Első szerző:Török Rebeka
Cím:Evaluation of Possible Processing Time Effects on the Global N-Glycosylation Profile of Human Blood Samples / Rebeka Torok, Anna Farkas, András Guttman, Gabor Jarvas
Dátum:2020
ISSN:1566-5240
Megjegyzések:The utilization of N-glycan profiling recently gained high importance in fundamental biomedical and applied clinical research. However, for the time being, no glycan biomarker has been approved for clinical diagnosis by the regulatory agencies due to the lack of verifications on large patient cohorts and suitable analytical technologies. In this paper, the effect of human blood sample handling was studied prior to N-glycosylation profiling by capillary electrophoresis, coupled with high sensitivity fluorescence detection. Special attention was paid to the preservation of sialylated structures because of their important clinical - biological relevance. Our results suggested that it is adequate to refrigerate and store the collected total blood samples prior to analysis to obtain unbiased results. Furthermore, we report on the good practice of serum sample handling in order to prevent decomposition of the sialylated structures. Our findings may promote procedure standardization and easier clinical translation of diagnostic N-glycosylation profiling in molecular medicinal applications.
Tárgyszavak:Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban
folyóiratcikk
Megjelenés:Current Molecular Medicine. - 20 : 10 (2020), p. 840-846. -
További szerzők:Farkas Anna (1988-) (molekuláris biológus) Guttman András (1954-) (vegyészmérnök) Járvás Gábor (1982-) (vegyészmérnök)
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DOI
Intézményi repozitóriumban (DEA) tárolt változat
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