CZE of human alpha‐1‐acid glycoprotein for qualitative and quantitative comparison of samples from different pathological conditions.

Lacunza, Izaskun; Sanz, Jesus; Diez-Masa, Jose Carlos; Frutos, Mercedes de

doi:10.1002/elps.200790010

Cited by 5 publications

(9 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Both the concentration and composition of AGP can change with various diseases and clinical conditions [2,[4][5][6][7]. For instance, the normal concentration of AGP in serum ranges from 0.5 to 1.0 mg/mL, or 12 to 24 μM; however, this concentration can increase by up to tenfold in some clinical situations [2].…”

Section: Introductionmentioning

confidence: 99%

“…For instance, a decrease in the degree of glycan branching has been reported in patients with infection and systemic lupus erythematosus (SLE) [4]. An increase in α1-3 fucosylation has been noted in pancreatic cancer [5], and an increase in levels of AGP sialylation has been seen in ovarian cancer and lymphoma [6,7]. These changes have made AGP glycoforms and their related glycosylation patterns of interest as potential biomarkers for these and other disease states [4][5][6][7].…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Glycoform Analysis of Alpha1-Acid Glycoprotein by Capillary Electrophoresis Using Electrophoretic Injection

Zhang

Clarke

Hage³

2019

Methods in Molecular Biology

View full text Add to dashboard Cite

Human alpha 1-acid glycoprotein (AGP) is an acute phase glycoprotein that has a heterogeneous glycosylation pattern. This pattern can change in certain diseases, which has resulted in interest in using AGP glycoforms as potential biomarkers for these diseases. This report describes a method that uses capillary electrophoresis to characterize and analyze AGP glycoforms both in purified samples of AGP and in human serum. This method uses static and dynamic coatings of poly (ethylene oxide) that are applied to a silica capillary for separation of AGP glycoforms in the reversed-polarity mode of CE and in the presence of negligible electroosmotic flow. Electrophoretic injection is performed onto such capillaries by using a stationary stacking interface between the sample and running buffer. In addition, acidic precipitation and desalting are used to allow for the isolation and the analysis of AGP from only 65 μL of serum. Up to eleven AGP glycoform bands can be reproducibly separated by this method, with the difference in migration time between neighboring bands being 12-to almost 60-fold larger than the standard deviation for the migration time of any given band. A limit of detection down to about 2 nM per glycoform band can be obtained by this method for AGP in serum based on absorbance detection and without the need for further sample modification or labeling.

show abstract

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Glycoform Analysis of Alpha1-Acid Glycoprotein by Capillary Electrophoresis Using Electrophoretic Injection

Zhang

Clarke

Hage³

2019

Methods in Molecular Biology

View full text Add to dashboard Cite

show abstract

“…Os procedimentos analíticos mais utilizados para quantificação da α1-AGP, em soro ou plasma, são a imunoturbidimetria (LE COUTRE et al, 2002;COLOMBO et al, 2006;MESTRINER et al, 2007), a imunodifusão radial (ARNOLD; MEYERSON, 1990;CLAPPERTON et al, 2007;PALTRINIERI et al, 2007), a imunoeletroforese por afinidade cruzada (SLUZEWSKA et al, 1996) e a eletroforese capilar (LACUNZA, et al, 2006).…”

Section: Ca Ap Pí íT Tu Ul Lo O 1 1: Análise Da Alfa-1 Glicoproteínunclassified

“…Em pHs, ligeiramente, acima de 4,0 esta proteína ainda encontrase na forma ionizada, ou seja, negativamente carregada. A maioria dos métodos desenvolvidos por CE para separar esta proteína utiliza capilares revestidos internamente e/ou soluções tampão com baixo valor de pH, com um ou mais aditivos (KAKEHI et al, 2001;SEI et al, 2002;LACUNZA et al, 2006). Estes procedimentos permitem uma melhor solubilidade e redução das interações da proteína com a parede interna do capilar.…”

Section: Otimização Do Métodounclassified

See 1 more Smart Citation

Aplicações da eletroforese capilar na análise do biomarcadir alfa-1 glicoproteína ácida, no controle de qualidade do biofármaco interferon alfa 2a e na avaliação da estabilidade enantiosseletiva do fármaco isradipina

Aguiar¹

View full text Add to dashboard Cite

pelo amor, carinho, alegrias, ensinamentos e educação, que sempre servirão como modelos de dedicação e trabalho. Aos meus irmãos, Rafael e Renato Rafael e Renato Rafael e Renato Rafael e Renato, pelo amor, diálogos, discussões, alegrias e tudo mais.... À Gabrie Gabrie Gabrie Gabriela la la la, pelo amor, carinho, diálogos, incentivos e companheirismo. Agradecimentos Agradecimentos Agradecimentos Agradecimentos Agradeço a Deus Deus Deus Deus pela vida, pela saúde, pelos momentos difíceis que nos ensinam e os alegres que nos revigoram. Aos meus pais Ademar Ademar Ademar Ademar e Elza Elza Elza Elza pela vida, carinho, incentivo e esforço para me proporcionar a melhor educação possível. Aos meus irmãos Rafael Rafael Rafael Rafael e Renato Renato Renato Renato, meus melhores amigos. Eu não teria conseguido estar onde estou se vocês não existissem. À Gabriela Gabriela Gabriela Gabriela pelo carinho e vida compartilhada nestes anos. Por incentivar e compartilhar todos os momentos da minha vida e principalmente desse projeto com dedicação e amor.

show abstract

Development and evaluation of silica-based lectin microcolumns for glycoform analysis of alpha1-acid glycoprotein

Zhang

Hage

2019

Analytica Chimica Acta

View full text Add to dashboard Cite

Silica-based lectin microcolumns were developed and optimized for the separation and analysis of glycoform fractions in alpha 1 -acid glycoprotein (AGP) based on both the degree of branching and level of fucosylation. Concanavalin A (Con A) and Aleuria Aurantia lectin (AAL) were immobilized onto HPLC-grade silica by reductive amination and packed into 2.1 mm i.d. × 5.0 cm microcolumns. Factors examined for these microcolumns include their protein content, binding capacity, binding strength and band-broadening under isocratic conditions (Con A) or step elution conditions (AAL) and in the presence of various flow rates or temperatures. These factors were examined by using experiments based on frontal analysis, zonal elution, peak profiling and peak decay analysis. Up to 200 μg AGP could be loaded onto a Con A microcolumn and provide linear elution conditions, and 100 μg AGP could be applied to an AAL microcolumn. The final conditions for separating retained and non-retained AGP glycoform fractions on a Con A microcolumn used a flow rate of 50 μL min −1 and a temperature of 50 °C, which gave a separation of these fractions within 20 min or less. The final conditions for an AAL microcolumn included a flow rate of 0.75 mL min −1 , a temperature of 50 °C, and the use of 2.0 mM L-fucose as a competing agent for elution, giving a separation of non-retained and retained AGP glycoforms in 6 min or less. The inter-day precisions were ± 0.7-4.0% or less for the retention times of the AGP glycoforms and ± 2.2-3.0% or less for their peak areas.

show abstract

CZE of human alpha‐1‐acid glycoprotein for qualitative and quantitative comparison of samples from different pathological conditions.

Cited by 5 publications

References 0 publications

Glycoform Analysis of Alpha1-Acid Glycoprotein by Capillary Electrophoresis Using Electrophoretic Injection

Glycoform Analysis of Alpha1-Acid Glycoprotein by Capillary Electrophoresis Using Electrophoretic Injection

Aplicações da eletroforese capilar na análise do biomarcadir alfa-1 glicoproteína ácida, no controle de qualidade do biofármaco interferon alfa 2a e na avaliação da estabilidade enantiosseletiva do fármaco isradipina

Development and evaluation of silica-based lectin microcolumns for glycoform analysis of alpha1-acid glycoprotein

Contact Info

Product

Resources

About