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1.

001-es BibID:BIBFORM065042
035-os BibID:(WoS)000365931100006 (Scopus)84948451094
Első szerző:Guttman András (vegyészmérnök)
Cím:Effect of Separation Temperature on Structure Specific Glycan Migration in Capillary Electrophoresis / Andras Guttman, Marta Kerekgyarto, Gabor Jarvas
Dátum:2015
ISSN:0003-2700
Megjegyzések:Temperature dependent differential migration shifts were studied in capillary electrophoresis between linear (maltooligosaccharides) and branched (sialylated, neutral and core fucosylated biantennary IgG glycans) carbohydrates. Background electrolytes without as well as with low and high molecular weight additives (ethylene glycol, linear polyacrylamide and poly(ethylene oxide)) were investigated for this phenomena in the temperature range of 20-50 ?C. Glucose unit (GU) value shifts were observed with increasing temperature for the all IgG glycans both in additive-free and additive-containing background electrolytes, emphasizing the importance of tight temperature control during glycosylation analysis by capillary electrophoresis. The activation energy concept was applied to understand the structure specific electrophoretic migration of the different sugar molecules. Activation energy values were derived from the slopes of the Arrhenius plots of logarithmic mobility vs reciprocal absolute temperature and compared for the linear and branched sugars as well as for the various background electrolyte additives.
Tárgyszavak:Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban
folyóiratcikk
capillary electrophoresis
glycan structure
mobility
temperature
activation energy
Megjelenés:Analytical Chemistry. - 87 : 23 (2015), p. 11630-11634. -
További szerzők:Kerékgyártó Márta (1987-) (molekuláris biológus) Járvás Gábor (1982-) (vegyészmérnök)
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2.

001-es BibID:BIBFORM065045
035-os BibID:(Scopus)84987657447 (WOS)000383596800009
Első szerző:Járvás Gábor (vegyészmérnök)
Cím:On the electromigration of charged-fluorophore labeled oligosaccharides in polyethylene oxide solutions / Gabor Jarvas, Marta Kerekgyarto, Andras Guttman
Dátum:2016
ISSN:0173-0835
Megjegyzések:The separation mechanism of charged-fluorophore (aminopyrenetrisulfonate) labeled maltooligosaccharides with ?1-4 linkage was studied in polyethylene oxide solutions (MW 300,000 Da) with special interest to possible analyte and/or network deformations as well as potential solute-matrix interactions. The electrophoretic mobilities of the APTS labeled maltooligosaccharides were found proportional with their MW-2/3 . The Arrhenius function was used to determine the activation energy needed by the labeled sugars to migrate through the separation media. With increasing solute size, the activation energy (Ea ) values decreased in polymer concentrations above the entanglement threshold of the polyethylene oxide, while showed apparently independent function at the entanglement threshold. The observed phenomenon was considered as a result of solute-matrix interaction, which could be alleviated by the addition of an organic modifier to the background electrolyte.
Tárgyszavak:Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban
folyóiratcikk
polymer network
polyethylene oxide
capillary electrophoresis
Megjelenés:Electrophoresis. - 37 : 17-18 (2016), p. 2347-2351. -
További szerzők:Kerékgyártó Márta (1987-) (molekuláris biológus) Guttman András (1954-) (vegyészmérnök)
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3.

001-es BibID:BIBFORM065038
Első szerző:Kerékgyártó Márta (molekuláris biológus)
Cím:Towards the generation of an aminonaphthalene trisulfonate labeled N-glycan database for capillary gel electrophoresis analysis of carbohydrates / Márta Kerékgyártó, András Guttman
Dátum:2014
ISSN:0173-0835
Megjegyzések:There is an increasing trend to develop therapeutic glycoproteins, mostly antibodies that require high resolution bioanalytical tools to address the challenging aspects of comprehensive carbohydrate characterization. In this paper we introduce an initial version of a glucose unit database for 8-aminonaphthalene-1,3,6-trisulfonic acid-labeled glycans. At this stage we mainly focused on therapeutic IgG derived glycans of core fucosylated biantennary structures with and without sialic acid residues, as well as high mannose structures. Currently 25 oligosaccharides represent this first set of the database that shows the abbreviated names of the individual sugar structures with their graphic representation, precise molecular mass and glucose unit (GU) values with corresponding SDs. The database will provide a quick glycan analysis tool for preliminary data interpretation of rapid (around 200 s) CGE-LED-induced fluorescence (CGE-LEDIF) based glycan profiling runs.
Tárgyszavak:Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban
glycan analysis
capillary gel electrophoresis
ANTS labeling
structural elucidation
database
Megjelenés:Electrophoresis. - 35 : 15 (2014), p. 2222-2228. -
További szerzők:Guttman András (1954-) (vegyészmérnök)
Pályázati támogatás:MTA-PE Translation Glycomics project
MTA
TÁMOP-4.2.4.A/2-11/1-2012-0001
TÁMOP
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4.

001-es BibID:BIBFORM065044
035-os BibID:(Scopus)84958616655 (WOS)000370657900002
Első szerző:Kerékgyártó Márta (molekuláris biológus)
Cím:Activation energy associated with the electromigration of oligosaccharides through viscosity modifier and polymeric additive containing background electrolytes / Márta Kerékgyártó, Gábor Járvás, Levente Novák, András Guttman
Dátum:2016
ISSN:0173-0835
Megjegyzések:The activation energy related to the electromigration of oligosaccharides can be determined from their measured electrophoretic mobilities at different temperatures. The effects of a viscosity modifier (ethylene glycol) and a polymeric additive (linear polyacrylamide) on the electrophoretic mobility of linear sugar oligomers with ?1-4 linked glucose units (maltooligosaccharides) were studied in CE using the activation energy concept. The electrophoretic separations of 8-aminopyrene-1,3,6-trisulfonate-labeled maltooligosaccharides were monitored by LIF detection in the temperature range of 20-50?C, using either 0-60% ethylene glycol (viscosity modifier) or 0-3% linear polyacrylamide (polymeric additive) containing BGEs. Activation energy curves were constructed based on the slopes of the Arrhenius plots. With the use of linear polyacrylamide additive, solute size-dependent activation energy variations were found for the maltooligosaccharides with polymerization degrees below and above maltoheptaose (DP 7), probably due to molecular conformation changes and possible matrix interaction effects.
Tárgyszavak:Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban
folyóiratcikk
capillary electrophoresis
activation energy
viscosity
Polyacrylamide
Megjelenés:Electrophoresis. - 37 : 4 (2016), p. 573-578. -
További szerzők:Járvás Gábor (1982-) (vegyészmérnök) Novák Levente (1967-) (biológus) Guttman András (1954-) (vegyészmérnök)
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5.

001-es BibID:BIBFORM065046
Első szerző:Kerékgyártó Márta (molekuláris biológus)
Cím:Capillary Gel Electrophoresis / Márta Kerékgyártó, András Guttman
Dátum:2015
Megjegyzések:Capillary gel electrophoresis is a rapidly developing technique, which has been successfully used in the bioanalytical field, especially for separation of biopolymers like nucleic acids, proteins, and complex carbohydrates. This chapter starts with a summary of the theoretical and practical aspects of capillary gel electrophoresis including separation principles, instrument design, capillary coating techniques, and the most frequently used sieving matrices. It is followed by the description of the various detection options, such as UV-Vis, diode-array, laser-induced fluorescence, and light-emitting diode-induced fluorescence types. Finally, an overview of the key application areas is given with the main emphasis on nucleic acid, protein, and complex carbohydrate analysis, as well as the latest developments in microchip electrophoresis devices.
Tárgyszavak:Orvostudományok Elméleti orvostudományok könyvfejezet
carbohydrates
capillary gel electrophoresis
lab-on-a-chip
nucleic acids
proteins
sieving matrices
Megjelenés:Analytical Separation Science / Ed. Jared L. Anderson, Alain Berthod, Verónica Pino Estévez, Apryll M. Stalcup. - p. 555-580
További szerzők:Guttman András (1954-) (vegyészmérnök)
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6.

001-es BibID:BIBFORM050944
Első szerző:Kerékgyártó Márta (molekuláris biológus)
Cím:Neoglycoproteins as carbohydrate antigens : synthesis, analysis, and polyclonal antibody response / Márta Kerékgyártó, Anikó Fekete, Zoltán Szurmai, János Kerékgyártó, László Takács, István Kurucz, András Guttman
Dátum:2013
ISSN:0173-0835
Megjegyzések:The analysis and polyclonal antibody response for newly synthesized maltose-BSA conjugate neoglycoproteins is described. In this first proof of concept study, a simple carbohydrate antigen, maltose, was linked to BSA by reductive amination. An aglycone spacer was utilized to conserve the intact annular maltose structure and to promote the accessibility of the carbohydrate immunogen hapten during immunization. The neoglycoproteins were investigated by CGE and the number of conjugated maltose residues was determined by MALDI-TOF MS. The neoglycoproteins were then evaluated by immunization of BALB/c mice and the polyclonal antibody response was tested by ELISA as evidence for the presence of sugar-containing epitope-specific antibodies. Selective antibody binding was demonstrated to the synthesized neoglycoproteins with different (low and high) glycosylation degrees suggesting the possible use of this approach to generate antibodies. Moreover, the polyclonal antibody response was not inhibited by maltose or other simple carbohydrates to confirm presence of the neoglycoprotein-specific antibodies.
Tárgyszavak:Természettudományok Kémiai tudományok idegen nyelvű folyóiratközlemény külföldi lapban
Carbohydrate antigen
Carbohydrate-specific antibody
ELISA
Neoglyco-protein
Megjelenés:Electrophoresis. - 34 : 16 (2013), p. 2379-2386. -
További szerzők:Fekete Anikó (1973-) (vegyész) Szurmai Zoltán (1953-) (szerves kémikus) Kerékgyártó János (1957-) (biokémikus, vegyész) Takács László (1955-) Kurucz István Guttman András (1954-) (vegyészmérnök)
Pályázati támogatás:K-81839
OTKA
TÁMOP-4.2.2.A-11/1/KONV-2012-0023-"VÉD-ELEM"
TÁMOP
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7.

001-es BibID:BIBFORM050946
Első szerző:Kerékgyártó Márta (molekuláris biológus)
Cím:Ultrafast haplotyping of putative microRNA-binding sites in the WFS1 gene by multiplex polymerase chain reaction and capillary gel electrophoresis / Márta Kerékgyártó, Nóra Németh, Tamás Kerekes, Zsolt Rónai, András Guttman
Dátum:2013
ISSN:0021-9673
Megjegyzések:The transmembrane protein wolframin (WSF1) plays a crucial role in cell integrity in pancreatic beta cells and maintaining ER homeostasis. Genetic variations in the WFS1 gene have been described to be associated with Wolfram syndrome or type 2 diabetes mellitus. In this paper we report on an efficient double-tube allele-specific amplification method in conjunction with ultrafast capillary gel electrophoresis for direct haplotyping analysis of the SNPs in two important miRNA-binding sites (rs1046322 and rs9457) in the WFS1 gene. An automated single-channel capillary gel electrophoresis system was utilized in the method that provided dsDNA fragment analysis in less than 240 s. The light-emitting diode induced fluorescence (LEDIF) detection system enabled excellent sensitivity for automated haplotyping of a large number of clinical samples. The detection limit was 0.002 ng/?L using field amplified injection from water diluted samples. The dynamic quantitation range was 0.08?10.00 ng/?L (R2 = 0.9997) in buffer diluted samples.
Tárgyszavak:Természettudományok Kémiai tudományok idegen nyelvű folyóiratközlemény külföldi lapban
WFS1 gene
miRNA-binding site
Haplotype
SNP
Double-tube allele-specific PCR
Capillary gel electrophoresis
Megjelenés:Journal of Chromatography A. - 1286 (2013), p. 229-234. -
További szerzők:Németh Nóra Kerekes Tamás Rónai Zsolt Guttman András (1954-) (vegyészmérnök)
Pályázati támogatás:K-81839
OTKA
K83766
OTKA
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8.

001-es BibID:BIBFORM050949
Első szerző:Kerékgyártó Márta (molekuláris biológus)
Cím:Light-emitting diode induced fluorescence (LED-IF) detection design for a pen-shaped cartridge based single capillary electrophoresis system / Márta Kerékgyártó, Tamás Kerekes, Eric Tsai, Varoujan D. Amirkhanian, András Guttman
Dátum:2012
ISSN:0173-0835
Megjegyzések:CGE is a well-established separation technique for the analysis of biologically important molecules such as nucleic acids. The inherent high resolving power, rapid analysis times, excellent detection sensitivity, and quantification capabilities makes this method favorable compared to conventional manual polyacrylamide and agarose slab gel electrophoresis techniques. In this paper we introduce a novel single-channel capillary gel electrophoresis system with LED-induced fluorescence detection also utilizing a compact pen-shaped capillary cartridge design for automatic analysis of samples from a 96-well plate. To evaluate the suitability of the system, 1000 genomic DNA(gDNA) samples were analyzed in gel filled capillaries and detected by the microball ended excitation and emission optical fiber based LED-induced fluorescence detection system. Excellent migration time reproducibility of RSD <0.75% was obtained over the course of 1000 runs. The system rapidly distinguished between intact and degraded gDNA samples, therefore provided important information if they could be used for downstream quantitative PCR processing where high-quality intact gDNA was key. We envision that this novel system design will rapidly find new applications in both research and clinical diagnostic laboratories as a highly sensitive and easy to use bio-analytical approach.
Tárgyszavak:Természettudományok Kémiai tudományok idegen nyelvű folyóiratközlemény külföldi lapban
Fluorescence detection
Genomic DNA purity
Light emitting diode
Quality control
Single-channel capillary gel electrophoresis
Megjelenés:Electrophoresis. - 33 : 17 (2012), p. 2752-2758. -
További szerzők:Kerekes Tamás Tsai, Eric Amirkhanian, Varoujan D. Guttman András (1954-) (vegyészmérnök)
Pályázati támogatás:K-81839
OTKA
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9.

001-es BibID:BIBFORM065037
Első szerző:Németh Nóra
Cím:Rapid identification of human SNAP-25 transcript variants by a miniaturized capillary electrophoresis system / Nóra Németh, Márta Kerékgyártó, Mária Sasvári-Székely, Zsolt Rónai, András Guttman
Dátum:2014
ISSN:0173-0835
Megjegyzések:The 25 kDa synaptosomal-associated protein (SNAP-25) is a crucial component of the soluble N-ethylmaleimide-sensitive factor attachment protein receptor complex and plays an important role in neurotransmission in the central nervous system. SNAP-25 has two different splice variants, SNAP-25a and SNAP-25b, differing in nine amino acids that results in a slight functional alteration of the generated soluble N-ethylmaleimide-sensitive factor attachment protein receptor complex. Two independent techniques, a PCR-miniaturized CE method and a real-time PCR based approach were elaborated for the specific and quantitative detection of the two SNAP-25 transcription variants. DNA-constructs coding for the two isoforms were used for optimization. Excellent specificity was observed with the use of our previously described highly sensitive miniaturized CE system in combination with quantitative PCR. The ratio of the two isoforms were reliably detected in a range of at least four orders of magnitude with a linear regression of R(2) = 0.987. Expression of the two isoforms was determined in human samples, where SNAP-25 was detected even in non-neural tissues, although at approximately a 100-fold lower level compared to the central nervous system. The relative amount of the SNAP-25b isoform was higher in the brain, whereas expression of SNAP-25a variant proved to be slightly higher in extra-neural cell types. The genomics approach in conjunction with the miniaturized CE system introduced in this paper is readily applicable for rapid alternative splice variant analysis.
Tárgyszavak:Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban
SNAP-25
transcription variant
real-time PCR
restriction endonuclease
capillary electrophoresis
Megjelenés:Electrophoresis. - 35 : 2-3 (2014), p. 379-384. -
További szerzők:Kerékgyártó Márta (1987-) (molekuláris biológus) Sasvári-Székely Mária Rónai Zsolt Guttman András (1954-) (vegyészmérnök)
Pályázati támogatás:MTA-PE Translational Glycomics project
MTA
K81839
OTKA
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10.

001-es BibID:BIBFORM065041
Első szerző:Székely Andrea
Cím:Multi Capillary SDS-Gel Electrophoresis for the Analysis of Fluorescently Labeled mAb preparations: a high throughput quality control process for the production of QuantiPlasma and PlasmaScan mAb libraries / Andrea Székely, Ákos Szekrényes, Márta Kerékgyártó, Attila Balogh, János Kádas, József Lázár, András Guttman, István Kurucz, László Takács
Dátum:2014
ISSN:0173-0835
Megjegyzések:Molecular heterogeneity of mAb preparations is the result of various co- and post-translational modifications and to contaminants related to the production process. Changes in molecular composition results in alterations of functional performance, therefore quality control and validation of therapeutic or diagnostic protein products is essential. A special case is the consistent production of mAb libraries (QuantiPlasma? and PlasmaScan?) for proteome profiling, quality control of which represents a challenge because of high number of mAbs (>1000). Here, we devise a generally applicable multicapillary SDS-gel electrophoresis process for the analysis of fluorescently labeled mAb preparations for the high throughput quality control of mAbs of the QuantiPlasma? and PlasmaScan? libraries.
Tárgyszavak:Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban
capillary electrophoresis
glycoform
mAb proteomics
quality control
SDS
Megjelenés:Electrophoresis. - 35 : 15 (2014), p. 2155-2162. -
További szerzők:Szekrényes Ákos (1983-) (vegyészmérnök) Kerékgyártó Márta (1987-) (molekuláris biológus) Balogh Attila Kádas János (1976-) (molekuláris biológus, biokémikus, kertészmérnök) Lázár József Guttman András (1954-) (vegyészmérnök) Kurucz István Takács László (1955-)
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11.

001-es BibID:BIBFORM050948
Első szerző:Szekrényes Ákos (vegyészmérnök)
Cím:High-throughput analysis of therapeutic and diagnostic monoclonal antibodies by multicapillary SDS gel electrophoresis in conjunction with covalent fluorescent labeling / Ákos Szekrényes, Udo Roth, Márta Kerékgyártó, Andrea Székely, István Kurucz, Karen Kowalewski, András Guttman
Dátum:2012
ISSN:1618-2642
Megjegyzések:Capillary gel electrophoresis (CGE) in the presence of sodium dodecyl sulfate (SDS) is a well-established and widely used protein analysis technique in the biotechnology industry, and increasingly becoming the method of choice that meets the requirements of the standards of International Conference of Harmonization (ICH). Automated single channel capillary electrophoresis systems are usually equipped with UV absorbance and/or laser-induced fluorescent (LIF) detection options offering general applicability and high detection sensitivity, respectively; however, with limited throughput. This shortcoming is addressed by the use of multicapillary gel electrophoresis (mCGE) systems with LED-induced fluorescent detection (LED-IF), also featuring automation and excellent detection sensitivity, thus widely applicable to rapid and large-scale analysis of biotherapeutics, especially monoclonal antibodies (mAb). The methodology we report in this paper is readily applicable for rapid purity assessment and subunit characterization of IgG molecules including detection of non-glycosylated heavy chains (NGHC) and separation of possible subunit variations such as truncated light chains (Pre-LC) or alternative splice variants. Covalent fluorophore derivatization and the mCGE analysis of the labeled IgG samples with multi-capillary gel electrophoresis are thoroughly described. Reducing and non-reducing conditions were both applied with and without peptide N-glycosidase F mediated deglycosylation.
Tárgyszavak:Természettudományok Kémiai tudományok idegen nyelvű folyóiratközlemény külföldi lapban
Biotherapeutics
Quality analysis
Multicapillary gel electrophoresis
Antibody purity
Fluorescent labeling
Megjelenés:Analytical and Bioanalytical Chemistry. - 404 : 5 (2012), p. 1485-1494. -
További szerzők:Roth, Udo Kerékgyártó Márta (1987-) (molekuláris biológus) Székely Andrea Kurucz István Kowalewski, Karen Guttman András (1954-) (vegyészmérnök)
Pályázati támogatás:K-81839
OTKA
TECH-09-A1-2009-0113
Egyéb
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