The present invention relates to a method for in vitro creation of
molecular libraries evolution of protein function. Particularly, it
relates to variability and modification of protein function by shuffling
polynucleotide sequence segments. A protein of desired characteristics
can be obtained by incorporating variant peptide regions (variant motifs)
into defined peptide regions (scaffold sequence). The variant motifs can
be obtained from parent DNA which has been subjected to mutagenesis to
create a plurality of differently mutated derivatives thereof or they can
be obtained from in vivo sequences. These variant motifs can then be
incorporated into a scaffold sequence and the resulting coded protein
screened for desired characteristics. This method is ideally used for
obtaining antibodies with desired characteristics by isolating individual
CDR DNA sequences and incorporating them into a scaffold which may, for
example, be from a totally different antibody.