A novel monoclonal antibody targeting carboxymethyllysine, an advanced glycation end product in atherosclerosis and pancreatic cancer

Abstract

Advanced glycation end products are formed by non-enzymatic reactions between proteins and carbohydrates, causing irreversible lysine and arginine alterations that severely affect protein structure and function. The resulting modifications induce inflammation by binding to scavenger receptors. An increase in advanced glycation end products is observed in a number of diseases e.g. atherosclerosis and cancer. Since advanced glycation end products also are present in healthy individuals, their detection and quantification are of great importance for usage as potential biomarkers. Current methods for advanced glycation end product detection are though limited and solely measure total glycation. This study describes a new epitope-mapped single chain variable fragment, D1-B2, against carboxymethyllysine, produced from a phage library that was constructed from mouse immunizations. The phage library was selected against advanced glycation end product targets using a phage display platform. Characterization of its binding pattern was performed using large synthetic glycated peptide and protein libraries displayed on microarray slides. D1-B2 showed a preference for an aspartic acid, three positions N-terminally from a carboxymethyllysine residue and also bound to a broad collection of glycated proteins. Positive immunohistochemical staining of mouse atherosclerotic plaques and of a tissue microarray of human pancreatic tumors confirmed the usability of the new scFv for advanced glycation end product detection in tissues. This study demonstrates a promising methodology for high-throughput generation of epitope-mapped monoclonal antibodies against AGE.

Receptor of advanced glycation end-products axis and gallbladder cancer: A forgotten connection that we should reconsider

“…Recent evidence has also revealed that the antibody targeting of RAGE ligands such as HMGB1 and AGEs may effectively decrease tumor progression in solid malignancies[ 74 ]. This approach can even enhance the antitumoral response of cancer immunotherapies by remobilizing the antitumor immune response[ 75 ].…”

Importance of Advanced Glycation End Products (AGE) in Human Disease and Diagnosis

“…AGEs have been observed not only in diseases such as atherosclerosis, rheumatoid arthritis, Alzheimer's disease and diabetes but various forms of cancer [35]. People with diabetes are at a significantly higher risk of many forms of cancer according to epidemiologic evidence.…”

Advanced Glycation End Products (AGEs): Biochemistry, Signaling, Analytical Methods, and Epigenetic Effects

“…However, methods based on the detection of autofluorescence of AGEs are limited as they exclusively measure the total fluorescent glycation. New promising methodologies for high-throughput generation of monoclonal antibodies mapped with epitopes against AGE have been applied and validated [26,27]. Signs of progress have been obtained in identifying epitopes, reaching a high level of sensitivity and a simple analytical procedure [26,27].…”

“…New promising methodologies for high-throughput generation of monoclonal antibodies mapped with epitopes against AGE have been applied and validated [26,27]. Signs of progress have been obtained in identifying epitopes, reaching a high level of sensitivity and a simple analytical procedure [26,27]. However, improvements in cultivation techniques and in the construction of specific monoclonal epitopes represent important goals to be achieved.…”

“…In recent years, new promising methodologies for highthroughput generation of monoclonal antibodies mapped with epitopes against AGE have been applied and validated. For example, recent studies have allowed the generation of new monoclonal antibodies destined for carboxymethyllysine [26,27]. Monoclonal antibodies have the potential to detect and quantify single glycated epitopes, but the effective and specific ones currently available are limited.…”