ADLM2025

Warmstart Modification of Enzymes with Recombinant Camelid VHH Domains

Tuesday, July 29
11:00 - 11:20 a.m.
Exhibit Hall Theater 3
Supported by Fortis Life

Overview

The use of isothermal amplification technology in molecular diagnostics has created a demand for warmstart modified enzymes that are inactive at ambient temperature but progressively activated as reactions are heated to temperatures above 50°C. While a variety of chemical and biological methods are available for hotstart modification of enzymes, such as DNA polymerases, these modalities typically require sustained temperatures >90°C for complete activation. Here we describe a novel workflow for the discovery, screening, and validation of recombinant camelid heavy-chain variable (VHH) domains, which can be used to warmstart modify enzymes used in isothermal amplification. As a proof-of-principle we describe the development of a novel warmstart VHH for Bst DNA polymerase that prevents off-target DNA amplification in a common LAMP assay format. This approach to enzyme warmstart modification presents a simple, yet robust, way to control reactions.

Speaker

Brendan Looyenga, PhD
Principal Scientist, Fortis Life

Sam Sugerman, PhD
Principal Scientist, Fortis Life