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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:BIBFORM085475
Első szerző:Járvás Gábor (vegyészmérnök)
Cím:Characterization of a Porous Nano-electrospray Capillary Emitter at Ultra-low Flow Rates / Jarvas Gabor, Fonslow Bryan, Yates John R., Foret Frantisek, Guttman Andras
Dátum:2017
ISSN:0021-9665
Megjegyzések:Biopharmaceuticals, especially therapeutic monoclonal antibodies, have emerged as a very promising new generation of protein-based drugs. However, their comprehensive analysis continues to pose new challenges for the bioanalytical field. Hyphenation of capillary electrophoresis with electrospray ionization (CE-MS) is a promising technique to address these challenges. One of the main advantages of CE-MS is the ability to produce stable electrospray at ultra-low flow rates (5?20 nl/min range). In this short communication we report on the characterization of a porous nanoelectrospray capillary emitter focusing on the effects of ultra-low flow rate on ionization efficiency, ion suppression and detection sensitivity. Ion suppression effect of a poorly-ionizable sugar in the presence of an easily-ionized peptide was reduced by almost 2-fold. Intact therapeutic antibody infusion analysis demonstrated that MS detection sensitivity increased by an order of magnitude with the decrease of flow rate from 250 nL/min to 20 nL/min using the nano-electrospray capillary emitter.
Tárgyszavak:Orvostudományok Klinikai orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban
folyóiratcikk
Megjelenés:Journal Of Chromatographic Science. - 55 : 1 (2017), p. 47-51. -
További szerzők:Fonslow, Bryan Yates, John R. Foret, František Guttman András (1954-) (vegyészmérnök)
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