VHH characterization. Comparison of recombinant with chemically synthesized anti‐HER2 VHH

• DOI: 10.1002/pro.3712
• Publication Date: 2019-08-19T08:25:58Z
• Authors: Lucie Hartmann; Thomas Botzanowski; Mathieu Galibert; Magali Jullian; Eric Chabrol; Gabrielle Zeder‐Lutz; Valérie Kugler; Johann Stojko; Jean‐Marc Strub; Gilles Ferry; Lukasz Frankiewicz; Karine Puget; Renaud Wagner; Sarah Cianférani; Jean A. Boutin
• Keywords: Protein Engineering

In the continuous exploration of VHH chemistry, biochemistry and therapeutic future use, we investigated two different production strategies this small antibody-like protein, using an anti-HER2 as a model. The total chemical synthesis 125 amino-acid peptide was performed with reasonable yield, even if optimization will be necessary to upgrade kind production. parallel, expressed same sequence in hosts: Escherichia coli Pichia pastoris. Both productions were successful led fair amount VHHs. integrity conformation characterized by complementary mass spectrometry approaches, while surface plasmon resonance experiments used assess recognition capacity affinity toward its "antigen." Using combination orthogonal techniques, it possible show that three VHHs-whether synthetic or recombinant ones-were properly similarly folded recognized "antigen" HER2 similar affinities, nanomolar range. This opens route further modified unnatural amino acids subsequently, VHH-drug conjugates.