Nature has successfully improved enzymes for many years by mutation and selection, but because of the long and slow nature of this process, many in vitro technologies have been set-up in order to mimic nature and evolve proteins rapidly.
All these technologies are unable to create a large diversity where all mutations (transitions and transversions) appear and are randomly spread along the gene sequence. MutaGen™ a new technology for molecular evolution developed by MilleGen is able to mimic nature quickly and produce a large diversity (patent WO 02387566; Mondon et al, Biotechnology J. 2007, 2:76-82; Emond et al, Protein Eng. Des. Sel. 2008, 21:267-274).
Based on an in vitro replication with low fidelity human DNA polymerases « mutases », MutaGen™ technology provides a powerful and simple approach to carrying out protein optimisation in the test tube.
It is a highly flexible, easy process, fast and usable for all proteins. The MutaGen™ technology allows to build up the largest diversity (up to 1010
variant molecules) for optimising recombinant proteins. In only one step, the number of variants could reach 90% or 100% with an average of mutation frequency from 3 to 15 mutations per kilobase. Furthermore, MutaGen takes advantages of error specificities of several error prone polymerases. These features show the innovating character of our proprietary technology in the directed evolution area.
Truly random mutations
During the mutation process, the mutase produces truly random misincorporation of nucleotides across the entire target DNA.
Controllable mutation rate
A controllable mutation rate with MutaGen™ provides useful nucleotide changes and increases the chances to isolate an improved protein. All type of mutations are created, transitions (65%) as well as transversions (35%).
Maximum diversity
MutaGen™ technology has the ability to sample a high range of protein variants in a single step. During one step replication of 1 microgram DNA more than 1010 DNA variants are made. The use of several polymerases allow the creation of libraries with different mutation rates, characteristic amount of the mutation subtypes and distinct mutational spectra.
Rapid and adaptable
Because of the high diversity generated in one single step of replication and because of the controlled mutation rate only few cycles of mutations and selection may be needed to produce a protein with the desired properties. MutaGen™ is also a technology, which generates a library of mutants, which can be easily screened.
