Advanced glycation end-products prepared in solution under high pressure contain epitopes distinct from those formed in the dry reaction at high temperature.
BACKGROUND
Advanced glycation end-products play an important role in diseases related to diabetes and aging processes. Model compounds are synthesized in order to prepare the diagnostic and experimental tools for studying the mechanisms of pathogenesis. The objective of the present study was to accelerate glycation and upgrade its efficiency under high-pressure conditions.


METHODS
Aqueous solutions of proteins were kept with carbohydrates under a pressure of up to 850 MPa for several hours. Then the high-pressure glycation (HPG) products were fractionated on a Sephadex G-200 column and characterized with SDS-PAGE and MALDI-TOF mass spectrometry.


RESULTS
The low-molecular-mass fraction of glycated proteins was separated from the two fractions containing high-and intermediate-molecular-mass cross-linked products of glycation. The products were then compared with those obtained with the high-temperature glycation (HTG) procedure carried out in dry conditions with a lyophilized mixture of substrates. The fractionated products were used to prepare rabbit sera.


CONCLUSIONS
The immunoblotting experiments showed that the epitopes on the cross-linked glycation products formed in solution under high pressure differed from those originating in dry conditions at high temperature. Sera against the HPG products were specific to homologous material and did not interact with the fractions obtained by HTG. The antibodies against HTG products recognized HTG but not HPG products.
Staniszewska
Magdalena
M
Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Wrocław, Poland.
Jarosz
Jerzy
J
Jon
Marek
M
Gamian
Andrzej
A
D000818:Animals; D000906:Antibodies; D002417:Cattle; D002850:Chromatography, Gel; D000939:Epitopes / Q000737:chemistry / Q000302:isolation & purification; D017127:Glycation End Products, Advanced / Q000737:chemistry* / Q000276:immunology* / Q000302:isolation & purification; D006023:Glycoproteins / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification; D006031:Glycosylation; D006358:Hot Temperature; D066298:In Vitro Techniques; D015416:Maillard Reaction; D008970:Molecular Weight; D011312:Pressure; D011817:Rabbits; D012710:Serum Albumin, Bovine / Q000737:chemistry / Q000276:immunology; D012996:Solutions; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D000906:Antibodies; D000939:Epitopes; D017127:Glycation End Products, Advanced; D006023:Glycoproteins; D012996:Solutions; D012710:Serum Albumin, Bovine