Capillary Zone Electrophoresis study of the in vitro albumin glycation process

Reducing sugars (such as glucose) can interact non-enzymatically with free amino-groups of proteins to form advanced glycation endproducts (AGEs). The objective of this work was to study the process of albumin glycation in vitro by Capillary Zone Electrophoresis (CZE). Bovine albumin (BSA; 50 mg/mL)...

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Autores principales: Castañon, María Mercedes, Steyerthal, Noemí Lucía, Castagnino, Juan Miguel, Garbosa, Graciela
Publicado: 2006
Materias:
Advanced glycation endproducts
Aminoguanidine
Bovine serum albumin
Capillary electrophoresis
Fluorometric analysis
Human serum albumin
Bovinae
Acceso en línea:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_03252957_v40_n4_p473_Castanon
http://hdl.handle.net/20.500.12110/paper_03252957_v40_n4_p473_Castanon
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Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires
id paper:paper_03252957_v40_n4_p473_Castanon
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spelling paper:paper_03252957_v40_n4_p473_Castanon2023-06-08T15:32:22Z Capillary Zone Electrophoresis study of the in vitro albumin glycation process Castañon, María Mercedes Steyerthal, Noemí Lucía Castagnino, Juan Miguel Garbosa, Graciela Advanced glycation endproducts Aminoguanidine Bovine serum albumin Capillary electrophoresis Fluorometric analysis Human serum albumin Bovinae Reducing sugars (such as glucose) can interact non-enzymatically with free amino-groups of proteins to form advanced glycation endproducts (AGEs). The objective of this work was to study the process of albumin glycation in vitro by Capillary Zone Electrophoresis (CZE). Bovine albumin (BSA; 50 mg/mL) and human serum (HS) were incubated with glucose (0.5 M) at 37°C for 12, 24 and 60 days. Aminoguanidine (AG) was used as inhibitor (0.5 M). Afterwards, HS-albumin was separated by precipitation and redissolved in phosphate-saline buffer. Fluorescent AGEs (λexcitation=338 nm, λ emission=442 nm) were detected after 12, 24 and 60 days of incubation. Fluorescence increased proportionally to the incubation times and AG was effective to inhibit the formation of fluorescent compounds. By CZE (boric acid 350 mM; pH 9,9; fused-silica gel capillary 50 μm x 50 cm; 25 kV; 90 μA; 20°C; λdetection=200 nm) longer migration times (MT) and wider peaks (PW-50) were observed, as the incubation time was prolonged. MT and PW-50 were partially reduced in the presence of AG. It can be concluded that CZE is a highly sensitive method to detect intermediate products during the protein glycation process. This procedure can be considered complementary to the fluorometric analysis used in AGEs' study. Fil:Castañón, M.M. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Steyerthal, N. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Castagnino, J.M. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Garbossa, G. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. 2006 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_03252957_v40_n4_p473_Castanon http://hdl.handle.net/20.500.12110/paper_03252957_v40_n4_p473_Castanon
institution Universidad de Buenos Aires
institution_str I-28
repository_str R-134
collection Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
topic Advanced glycation endproducts
Aminoguanidine
Bovine serum albumin
Capillary electrophoresis
Fluorometric analysis
Human serum albumin
Bovinae
spellingShingle Advanced glycation endproducts
Aminoguanidine
Bovine serum albumin
Capillary electrophoresis
Fluorometric analysis
Human serum albumin
Bovinae
Castañon, María Mercedes
Steyerthal, Noemí Lucía
Castagnino, Juan Miguel
Garbosa, Graciela
Capillary Zone Electrophoresis study of the in vitro albumin glycation process
topic_facet Advanced glycation endproducts
Aminoguanidine
Bovine serum albumin
Capillary electrophoresis
Fluorometric analysis
Human serum albumin
Bovinae
description Reducing sugars (such as glucose) can interact non-enzymatically with free amino-groups of proteins to form advanced glycation endproducts (AGEs). The objective of this work was to study the process of albumin glycation in vitro by Capillary Zone Electrophoresis (CZE). Bovine albumin (BSA; 50 mg/mL) and human serum (HS) were incubated with glucose (0.5 M) at 37°C for 12, 24 and 60 days. Aminoguanidine (AG) was used as inhibitor (0.5 M). Afterwards, HS-albumin was separated by precipitation and redissolved in phosphate-saline buffer. Fluorescent AGEs (λexcitation=338 nm, λ emission=442 nm) were detected after 12, 24 and 60 days of incubation. Fluorescence increased proportionally to the incubation times and AG was effective to inhibit the formation of fluorescent compounds. By CZE (boric acid 350 mM; pH 9,9; fused-silica gel capillary 50 μm x 50 cm; 25 kV; 90 μA; 20°C; λdetection=200 nm) longer migration times (MT) and wider peaks (PW-50) were observed, as the incubation time was prolonged. MT and PW-50 were partially reduced in the presence of AG. It can be concluded that CZE is a highly sensitive method to detect intermediate products during the protein glycation process. This procedure can be considered complementary to the fluorometric analysis used in AGEs' study.
author Castañon, María Mercedes
Steyerthal, Noemí Lucía
Castagnino, Juan Miguel
Garbosa, Graciela
author_facet Castañon, María Mercedes
Steyerthal, Noemí Lucía
Castagnino, Juan Miguel
Garbosa, Graciela
author_sort Castañon, María Mercedes
title Capillary Zone Electrophoresis study of the in vitro albumin glycation process
title_short Capillary Zone Electrophoresis study of the in vitro albumin glycation process
title_full Capillary Zone Electrophoresis study of the in vitro albumin glycation process
title_fullStr Capillary Zone Electrophoresis study of the in vitro albumin glycation process
title_full_unstemmed Capillary Zone Electrophoresis study of the in vitro albumin glycation process
title_sort capillary zone electrophoresis study of the in vitro albumin glycation process
publishDate 2006
url https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_03252957_v40_n4_p473_Castanon
http://hdl.handle.net/20.500.12110/paper_03252957_v40_n4_p473_Castanon
work_keys_str_mv AT castanonmariamercedes capillaryzoneelectrophoresisstudyoftheinvitroalbuminglycationprocess
AT steyerthalnoemilucia capillaryzoneelectrophoresisstudyoftheinvitroalbuminglycationprocess
AT castagninojuanmiguel capillaryzoneelectrophoresisstudyoftheinvitroalbuminglycationprocess
AT garbosagraciela capillaryzoneelectrophoresisstudyoftheinvitroalbuminglycationprocess
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