The intervening removable affinity tag (iRAT) production system facilitates Fv antibody fragment-mediated crystallography.

ABSTRACT: Fv antibody fragments have been used as co-crystallization partners in structural biology, particularly in membrane protein crystallography. However, there are inherent technical issues associated with the large-scale production of soluble, functional Fv fragments through conventional methods in various expression systems. To circumvent these problems, we developed a new method, in which a single synthetic polyprotein consisting of a variable light (V_L ) domain, an intervening removable affinity tag (iRAT), and a variable heavy (V_H ) domain is expressed by a Gram-positive bacterial secretion system. This method ensures stoichiometric expression of V_L and V_H from the monocistronic construct followed by proper folding and assembly of the two variab

SUBMITTER: Nomura Y 

PROVIDER: S-EPMC5119561 | biostudies-literature | 2016 Dec

REPOSITORIES: biostudies-literature