MethylTrap: A Supramolecular Affinity Column

Background

In proteomics and epigenetics, methyllysines act as ‘switches’ for gene expression and biological events, and have strong connections to disease biomarkers, treatment responses, and patient outcomes. Most methyllysines are low-abundance and difficult to identify and quantify in samples. Methyl-antibodies are continually noted for their poor performance in the lab. Most pan‑specific reagents that have been developed for methylysines are polycolonal antibodies that do not perform well and have significant batch‑to‑batch variability. Numerous papers have been published about their poor performance, outlining their lack of reliability, reproducibility, and pan-specificity. They are inherently inconsistent with no quality control.

Researchers at the University of Victoria (UVic) have developed the MethylTrap a supramolecular chemical affinity column for methylation‑targeted proteomics, featuring a range of compelling benefits.

Technology Overview

MethylTrap is a synthetic chemical affinity column for supramolecular sensing of methyllysines. When pan‑specificity is required the supramolecular sensing offered by MethylTrap has valuable advantages over biomolecular affinity agents. Supramolecular hosts have small binding surfaces that are perfect for binding individual functional groups while passing‑over surrounding structures on the target analyte. Methylysine interactions are increased by consistently grabbing more of the proteome allowing the methylated sites to be enriched; therefore, they are more likely to be detected in a standard proteomics analysis. MethylTrap is designed to be one simple enrichment step that slots right into the middle of what is otherwise a completely traditional proteomics workflow. There is a major advance in consistency when compared to polycolonal antibodies.

Benefits

 Scientific benefits:

• Unparalleled reliability and reproducibility, with superior day-to-day, batch-to-batch, and lab-to-lab reproducibility compared to antibodies.
• Pan-specificity and higher binding affinity, consistently revealing far more undiscovered mono/di/tri-methyllysine sites than found using antibodies
• Compatibility with existing liquid chromatography (LC-MS) setups

Commercial benefits:

• Manufacturability and stability: Reliable one-step synthesis of the chemical affinity handle and a longer, lower-maintenance shelf life than antibodies.
• Marketability: The chemical affinity handles can be bound to non-proprietary solid agarose supports and sold as columns, kits, or reagents.
• Expandability: New affinity handles can be developed to target different PTM residues

Applications

• Superior consistency and exploration of methylation sites and pathways in proteomics and epigenetics research, when compared to antibodies.
• Sample clean-up, separation, and enrichment before diagnostics with/without antibodies.
• Improved disease biomarker diagnostics

Opportunity