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001-es BibID:	BIBFORM106835
035-os BibID:	(Scopus)85138605879 (WoS)000856676100001
Első szerző:	Filep Csenge Boróka (biomérnök)
Cím:	Electromigration Dispersion in Sodium Dodecyl Sulfate Capillary Gel Electrophoresis of Proteins / Filep Csenge, Guttman András
Dátum:	2022
ISSN:	0003-2700
Megjegyzések:	The electromigration dispersion of the light- and heavy-chain subunit peaks of the therapeutic monoclonal antibody omalizumab was investigated in sodium dodecyl sulfate capillary gel electrophoresis (SDS?CGE) using borate cross-linked dextran sieving matrices. Increasing boric acid content (340?640 mM) caused electromigration dispersion shifts for both low (2%)- and high (10%)-dextran-concentration gels in all gel?buffer compositions. In case of the heavy-chain fragment, elevated borate concentrations resulted in decreasing tailing and increasing fronting with the use of higher- and lower-dextranconcentration gels, respectively. The light-chain fragment, on the other hand, exhibited increased fronting with increasing borate concentration for both dextran concentrations examined in this study. Increase of the glycerol ingredient level in the gel?buffer system caused the same effect as the increasing borate concentration in both dextran concentrations. The detected electromigration dispersion was considered as the result of the formation of monomeric and dimeric glycerol-borate complexes as co-ionic constituents, migrating slower than that of the unconjugated tetrahydroxyborate. In addition, complexation of the tetrahydroxyborate anion with the glucose building blocks of the dextran polymer decreased its mobility to practically zero, contributing to further decrease in the resultant effective mobility of the co-ionic species. We suggest that the observed fronting and/or tailing peak shapes of the monoclonal antibody fragments in SDS?CGE at increasing boric acid concentrations can be considered as the result of multiple effects including changes in pH, sieving matrix pore size, viscosity, and the mobility variation of the co-ionic borate adducts with the gel?buffer ingredients. While electromigration dispersion-mediated band broadening, in general, can be minimized via matching the effective mobility of the co-ionic species to the analyte molecules of interest, in case of borate cross-linked dextran gels, optimization of the boric acid concentration required special consideration of its gel cross-linking function. For the light- and heavy-chain fragments of the IgG analyte, best peak shapes were attained with the use of 10% dextran/340 mM boric acid and 10% dextran/640 mM boric acid-containing gel?buffer systems, respectively. Based on this observation, here we introduce the concept of borate-gradient-mediated transient mobility matching in SDS?CGE of proteins. This novel approach resulted in close to optimal peak shapes for the distantly migrating IgG subunits within a single run, as well as unraveled the long-sought possible solution to perform capillary pore-size-gradient gel electrophoresis.
Tárgyszavak:	Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban folyóiratcikk
Megjelenés:	Analytical Chemistry. - 94 : 38 (2022), p. 13092-13099. -
További szerzők:	Guttman András (1954-) (vegyészmérnök)
Pályázati támogatás:	EFOP-3.6.3-VEKOP-16-2017-00009 EFOP
Internet cím:	Intézményi repozitóriumban (DEA) tárolt változat DOI
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001-es BibID:	BIBFORM093742
035-os BibID:	(WoS)000623041000027 (Scopus)85100632127
Első szerző:	Filep Csenge Boróka (biomérnök)
Cím:	Effect of the Monomer Cross-Linker Ratio on the Separation Selectivity of Monoclonal Antibody Subunits in Sodium Dodecyl Sulfate Capillary Gel Electrophoresis / Csenge Filep, András Guttman
Dátum:	2021
ISSN:	0003-2700
Tárgyszavak:	Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban folyóiratcikk
Megjelenés:	Analytical Chemistry. - 93 : 7 (2021), p. 3535-3541. -
További szerzők:	Guttman András (1954-) (vegyészmérnök)
Pályázati támogatás:	2.3.2-15-2016-00017 GINOP NKFIH Egyéb ÚNKP-20-3-II-DE-294 Egyéb 2020-4.1.1-TKP2020 Egyéb
Internet cím:	Szerző által megadott URL DOI Intézményi repozitóriumban (DEA) tárolt változat
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001-es BibID:	BIBFORM083893
035-os BibID:	(WoS)000518234700070 (Scopus)85080047360
Első szerző:	Filep Csenge Boróka (biomérnök)
Cím:	The Effect of Temperature in Sodium Dodecyl Sulfate Capillary Gel Electrophoresis of Protein Therapeutics / Filep Csenge, Guttman András
Dátum:	2020
ISSN:	0003-2700
Tárgyszavak:	Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban folyóiratcikk
Megjelenés:	Analytical Chemistry. - 92 : 5 (2020), p. 4023-4028. -
További szerzők:	Guttman András (1954-) (vegyészmérnök)
Pályázati támogatás:	2.3.2-15-2016-00017 GINOP NKFIH NN 127062 Egyéb
Internet cím:	Szerző által megadott URL DOI Intézményi repozitóriumban (DEA) tárolt változat
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001-es BibID:	BIBFORM096076
035-os BibID:	(WoS)000672115800032 (Scopus)85110405663
Első szerző:	Guttman András (vegyészmérnök)
Cím:	Fundamentals of Capillary Electrophoretic Migration and Separation of SDS Proteins in Borate Cross-Linked Dextran Gels / András Guttman, Csenge Filep, Barry L. Karger
Dátum:	2021
ISSN:	0003-2700
Megjegyzések:	Abstract Abstract Image Recent progress in the development and production of new, innovative protein therapeutics require rapid and adjustable high-resolution bioseparation techniques. Sodium dodecyl sulfate capillary gel electrophoresis (SDS-CGE) using a borate (B) cross-linked dextran (D) separation matrix is widely employed today for rapid consistency analysis of therapeutic proteins in manufacturing and release testing. Transient borate cross-linking of the semirigid dextran polymer chains leads to a high-resolution separation gel for SDS?protein complexes. To understand the migration and separation basis of the D/B gel, the present work explores various gel formulations of dextran monomer (2, 5, 7.5, and 10%) and borate cross-linker (2 and 4%) concentrations. Ferguson plots were analyzed for a mixture of protein standards with molecular weights ranging from 20 to 225 kDa, and the resulting nonlinear concave curves pointed to nonclassical sieving behavior. While the 2% D/4% B gel resulted in the fastest analysis time, the 10% D/2% B gel was found to produce the greatest separation window, even higher than with the 10% D/4% B gel, due to a significant increase in the electroosmotic flow of the former composition in the direction opposite to SDS?protein complex migration. The study then focused on SDS-CGE separation of a therapeutic monoclonal antibody and its subunits. A combination of molecular weight and shape selectivity as well as, to a lesser extent, surface charge density differences (due to glycosylation on the heavy chain) influenced migration. Greater molecular weight selectivity occurred for the higher monomer concentration gels, while improved glycoselectivity was obtained using a more dilute gel, even as low as 2% D/2% B. This latter gel took advantage of the dextran?borate?glycoprotein complexation. The study revealed that by modulating the dextran (monomer) and borate (cross-linker) concentration ratios of the sieving matrix, one can optimize the separation for specific biopharmaceutical modalities with excellent column-to-column, run-to-run, and gel-to-gel migration time reproducibilities (<0.96% relative standard deviation (RSD)). The widely used 10% dextran/4% borate gel represents a good screening option, which can then be followed by a modified composition, optimized for a specific separation as necessary.
Tárgyszavak:	Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban folyóiratcikk
Megjelenés:	Analytical Chemistry. - 93 : 26 (2021), p. 9267-9276. -
További szerzők:	Filep Csenge Boróka (1993-) (biomérnök) Karger, Barry
Pályázati támogatás:	2.3.2- 15-2016-00017 GINOP NKFIH (NN 127062) Egyéb ÚNKP-20-3-II-DE-294 Egyéb 2020-4.1.1-TKP2020 Egyéb
Internet cím:	DOI Intézményi repozitóriumban (DEA) tárolt változat
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