Mutational analyses have been proved critical for determining the role of individual amino acids in proteins. Single amino acid changes can affect folding, thermodynamic stability, enzymatic activity, interactions, posttranslational modifications, and can even predict the severity of some diseases. Traditional methods to study the consequences of these changes are laborious. One must start by cloning the mutation, expressing and sometimes purifying the resulting variant, and finally testing its function in cellular or biochemical assays. Carrying out this approach on a protein of 100 amino acids would produce 1900 variants (100 amino acids × 19 changes), which cannot be all tested by traditional methods.
At the Multiplexed Assays Technology Hub (MATH), situated at The Walter and Eliza Hall Institute, we have been developing in house Deep Mutational Scanning protocols that measure the activity of 105 or more unique variants of a protein in a single experiment. This strategy provides unique insight into the sequence-function relationship of the proteins under study. The Multiplexed Assay Technology Hub can assist researchers by constructing libraries of variants, advising on screen design, and sequencing strategy, also performing bioinformatics analysis of the generated data.
Some applications that are suitable for Deep Mutational Scanning include predicting escape pathways during drug development, identifying antibody-antigen binding by epitope mapping, discrimination of disease-causing mutations from variants of unknown significance as well as many more.