Recent MIX-UP publication!

Different configurations of each new Nb compared to the structural model of the precursor protein.

Check out the new MIX-UP publication "ssDNA recombineering boosts in vivo evolution of nanobodies displayed on bacterial surfaces" published in the journal Communications Biology. Congratulations to our partners Yamal Al-ramahi, Akos Nyerges, Yago Margolles, Lidia Cerdán, Gyorgyi Ferenc, Csaba Pál, Luis Ángel Fernández and Víctor de Lorenzo to the success!

ssDNA recombineering has been exploited to hyperdiversify genomically-encoded nanobodies displayed on the surface of Escherichia coli for originating new binding properties. As a proof-of-principle a nanobody recognizing the antigen TirM from enterohaemorrhagic E. coli (EHEC) was evolved towards the otherwise not recognized TirM antigen from enteropathogenic E. coli (EPEC). To this end, E. coli cells displaying this nanobody fused to the intimin outer membrane-bound domain were subjected to multiple rounds of mutagenic oligonucleotide recombineering targeting the complementarity determining regions (CDRs) of the cognate VHH gene sequence. Binders to the EPEC-TirM were selected upon immunomagnetic capture of bacteria bearing active variants and nanobodies identified with a new ability to strongly bind the new antigen. The results highlight the power of combining evolutionary properties of bacteria in vivo with oligonucleotide synthesis in vitro for the sake of focusing diversification to specific segments of a gene (or protein thereof) of interest.

 

Click here to read the full article.