Antibody Fragments Humanization: Beginning with the End in Mind

International audienceMolecular engineering has made possible to reformat monoclonal antibodies into smaller antigen-binding structures like scFvs, diabodies, Fabs with new potential in vivo applications because they do not induce Fc-mediated functions. However, most of these molecules are from rodent origin. As a consequence, they are immunogenic and approval for administration to humans requires prior humanization. Today, there is no well-identified strategy to create recombinant humanized antibody V-domains that preserve the antigen-binding characteristics of the parental antibody associated with high stability and solubility. Here, we propose a strategy that consists in grafting CDRs onto properly chosen human antibody frameworks in order to reduce immunogenicity. A flowchart indicates the way to proceed in order to introduce an internal affinity purification tag while structural refinements are proposed to maintain antigen-binding characteristics. The best humanized candidates are identified through selection steps including in silico analysis, research scale production followed by early functional evaluation, purification assays, aggregation, and stability assessment

Influence of Zn and Mg Alloying on the Corrosion Resistance Properties of Al Coating Applied by Arc Thermal Spray Process in Simulated Weather Solution

In this study, Al–Zn and Al–Mg coatings were deposited on steel substrates by an arc thermal spray process. X-ray diffraction and scanning electr on microscopy were used to characterize the deposited coatings and corrosion products. Open circuit potential (OCP), electrochemical impedance spectroscopy, and potentiodynamic studies were used to assess the corrosion characteristics of these coatings after exposure according to the Society of Automotive Engineers (SAE) J2334 solution of varying durations. This solution simulates an industrial environment and contains chloride and carbonate ions that induce corrosion of the deposited coatings. However, the Al–Mg alloy coating maintained an OCP of approximately - 0.911 V versus Ag/AgCl in the SAE J2334 solution even after 792 h of exposure. This indicates that it protects the steel sacrificially, whereas the Al–Zn coating provides only barrier-type protection through the deposition of corrosion products. The Al–Mg coating acts as a self-healing coating and provides protection by forming Mg 6 Al 2 (OH) 16 CO 3 (Al–Mg layered double hydroxides). Mg 6 Al 2 (OH) 16 CO 3 has interlocking characteristics with a morphology of plate-like nanostructures and an ion-exchange ability that can improve the corrosion resistance properties of the coating. The presence of Zn in the corrosion products of the Al–Zn coating allows dissolution, but, at the same time, Zn 5 (OH) 6 (CO 3 ) 2 and Zn 6 Al 2 (OH) 16 CO 3 are formed and act to reduce the corrosion rate