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001-es BibID:BIBFORM113960
035-os BibID:(WoS)001043559400001 (Scopus)85166981215
Első szerző:Auer, Felicia
Cím:Size separation of sodium dodecyl sulfate-proteins by capillary electrophoresis in dilute and ultra-dilute dextran solutions / Felicia Auer, Andras Guttman
Dátum:2023
ISSN:0173-0835 1522-2683
Megjegyzések:SDS capillary gel electrophoresis is a widely used in the biopharma and the biomedical fields for rapid size separation of proteins. However, very limited information is available on the use of dilute and ultra-dilute sieving matrices for SDS?protein analysis. Here, background electrolytes (BGEs) containing 1%? 0% dextran were used in borate-based BGE to separate a protein sizing ladder (PSL) ?225 kDa and the intact and subunit forms of a therapeutic monoclonal antibody (mAb). The separation performance for the PSL and mAb components differed significantly with decreasing dextran concentration. Ferguson and reptation plots were used to elucidate the separation mechanism. Highly diluted dextran solutions resulted in linear Ferguson plots for both solute types (cf. Ogston theory) in spite of this model assumes a rigid pore structure, thus cannot describe the separation mechanism in ultra-dilute polymer solutions with no reticulations. The saddle differences between the resolution of the PSL and the intact/subunit mAb forms in ultra-dilute dextran-borate matrices suggested the importance of shape selectivity, manifested by the adequate separation of the SDS covered intact as well as light and heavy chain subunits of the therapeutic mAb even at zero dextran concentration.
Tárgyszavak:Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban
folyóiratcikk
borate
capillary gel electrophoresis
dextran
SDS-proteins
ultra-dilute polymer
Megjelenés:Electrophoresis. - 44 : 19-20 (2023), p. 1607-1614. -
További szerzők:Guttman András (1954-) (vegyészmérnök)
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2.

001-es BibID:BIBFORM099680
035-os BibID:(cikkazonosító)122497
Első szerző:Auer, Felicia
Cím:Recent advances in the analysis of human milk oligosaccharides by liquid phase separation methods / Auer Felicia, Jarvas Gabor, Guttman Andras
Dátum:2021
ISSN:1570-0232
Megjegyzések:Human milk is a complex, dynamically changing biological fluid, which contains a large amount of nonconjugated carbohydrates, referred to as human milk oligosaccharides (HMOs). These HMOs are very important for the infants as they play important roles in the formation of the gut microbiome, the immune system and support brain development. HMOs show highly complex structural diversity due to numerous linkage possibilities of the building monosaccharides. In order to elucidate their structure?function relationship and to develop more effective infant formulas, cutting-edge analytical technologies are in great demand. In this paper, we review the current strategies for HMO analysis based on liquid phase separation methods. High performance liquid chromatography, capillary electrophoresis and their hyphenation with mass spectrometry are critically reviewed, emphasizing their advantages and disadvantages from practical point of views. Recent advances of the methods are categorized according to their application fields.
Tárgyszavak:Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban
folyóiratcikk
Human milk
Oligosaccharides
Analysis
Chromatography
Electrophoresis
Mass spectrometry
Megjelenés:Journal Of Chromatography B-Analytical Technologies In The Biomedical And Life Sciences. - 1162 (2021), p. 1-9. -
További szerzők:Járvás Gábor (1982-) (vegyészmérnök) Guttman András (1954-) (vegyészmérnök)
Pályázati támogatás:2018-2.1.17-T?ET-KR-2018-00010
Egyéb
BIONANO_GINOP-2.3.2- 15-2016-00017
GINOP
NN127062
OTKA
TKP2020-IKA-07
OTKA
UNKP-20-5
Egyéb
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3.

001-es BibID:BIBFORM106825
035-os BibID:(cikkazonosító)8192 (scopus)85143597041 (wos)000896340300001
Első szerző:Torok Rebeka
Cím:The Effect of Sample Glucose Content on PNGase F-Mediated N-Glycan Release Analyzed by Capillary Electrophoresis / Torok Rebeka, Auer Felicia, Farsang Robert, Jona Eszter, Jarvas Gabor, Guttman Andras
Dátum:2022
ISSN:1420-3049
Megjegyzések:Protein therapeutics have recently gained high importance in general health care along with applied clinical research. Therefore, it is important to understand the structure?function relationship of these new generation drugs. Asparagine-bound carbohydrates represent an important critical quality attribute of therapeutic glycoproteins, reportedly impacting the efficacy, immunogenicity, clearance rate, stability, solubility, pharmacokinetics and mode of action of the product. In most instances, these linked N-glycans are analyzed in their unconjugated form after endoglycosidase-mediated release, e.g., PNGase F-mediated liberation. In this paper, first, N-glycan release kinetics were evaluated using our previously reported in-house produced 6His-PNGase F enzyme. The resulting deglycosylation products were quantified by sodium dodecyl sulfate capillary gel electrophoresis to determine the optimal digestion time. Next, the effect of sample glucose content was investigated as a potential endoglycosidase activity modifier. A comparative Michaelis-Menten kinetics study was performed between the 6His-PNGase F and a frequently employed commercial PNGase F product with and without the presence of glucose in the digestion reaction mixture. It was found that 1 mg/mL glucose in the sample activated the 6His-PNGase F enzyme, while did not affect the release efficiency of the commercial PNGase F. Capillary isoelectric focusing revealed subtle charge heterogeneity differences between the two endoglycosidases, manifested by the lack of extra acidic charge variants in the cIEF trace of the 6His-PNGase F enzyme, which might have possibly influenced the glucose-mediated enzyme activity differences.
Tárgyszavak:Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban
folyóiratcikk
Megjelenés:Molecules. - 27 : 23 (2022), p. 1-8. -
További szerzők:Auer, Felicia Farsang Róbert Jóna Eszter Járvás Gábor (1982-) (vegyészmérnök) Guttman András (1954-) (vegyészmérnök)
Pályázati támogatás:UNKP-22-3-I
Egyéb
Internet cím:Szerző által megadott URL
DOI
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