MutaMap™ Mutational Activity Map
high throughput service for mutational activity maps of therapeutic proteins
A new key tool for developability engineering
When it’s time to make final engineering decisions for your antibody or protein, MutaMap™ can help evaluate which individual point mutations to pursue. MutaMap™ is an in vitro assay system that helps explore the effect of substituting each amino acid in at each position in a protein sequence one by one with all 19 possible substitutions and find out the effect on protein activity. For example a sequence stretch of 100 amino acids will result in up to 2000 mutants to explore.
Mutations investigated in CDR1 and CDR3 of Avastin® heavy chain variable domain
For binding pair interactions MutaMap™ uses high throughput solution equilibrium titration (SET) immunoassays to determine the binding affinity for each construct tested.
MutaMap™ delivers a heat map for your protein (see Figure 1 above) that shows you which point
mutations lead to an increase, decrease, or no change in affinity (or
other activity) or non-function of the protein when interacting with one
or more of its binding partners. Effectively you can learn which
mutations, one by one, are likely to be permissible or favourable in your
protein in terms of the key property of binding to a ligand.
Focus on position S105 in CDR-H3 of Avastin® heavy chain variable domain
How does MutaMap™ compare to molecular evolution technologies?
Molecular evolution techniques such as phage display and other phenotype-genotype coupled randomization techniques are most commonly used in the affinity maturation process for monoclonal antibodies and other binding scaffolds. The advantage of these technologies is that they help explore a very large sequence space of combined mutations.
There comes a point however when final decisions have to be made on the implementation of a protein sequence where individual point mutations may be considered in an antibody or therapeutic protein to meet a variety of design objectives. Randomized molecular evolution is not appropriate for this step. Exploring individual point mutations is nothing new, but it has been difficult to carry this step out in very high throughput way, especially where the objective is to clone and express every mutant and then measure its affinity/activity with reasonable accuracy. This is what MutaMap™ can achieve.
Where does MutaMap™ fit in as part a project for e.g. generating a candidate monoclonal antibody for clinical development?
How long does a MutaMap™ project take?
Our objective is to complete medium size projects of exploring 500-2000 mutations in approximately 8-12 weeks from receiving the customer’s protein sequence. Larger projects will take slightly longer, depending on complexity.
What is the stepwise process for carrying out MutaMap™?
What will you get?
A final technical report delivered via our secure webserver showing you the affinity or activity determination for the wild type and each mutant with confidence interval. This will be presented in various formats for ease of interpretation, including a standard heatmap.
For customers that want to carry out protein antigenicity studies in parallel, these can be carried out in approximately the same timeframe as the MutaMap™. This means that within a period of approximately 8-12 weeks we will have determined experimentally both the putative T cell epitopes and the MutaMap™ of permitted mutations in your protein sequence. This information can allow you to proceed with much better informed decisions on how to address immunogenicity related issues for your program while addressing simultaneously other developability related design decisions for your sequence.