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Validation of the Production of Antibodies in Different Formats in the HEK 293 Transient Gene Expression System

Methods Mol Biol. 2021; 
Jens König, Michael Hust, Joop van den Heuvel
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PCR Cloning and Subcloning … pUC57-His8-proKLK-3C-hKLK7 (Cloning vector obtained from GenScript carrying the synthetic codon optimized DNA sequence His8 … maintenance of the columns and chromatography systems were done according to the protocols recommended by the manufacturing company … Get A Quote

摘要

Mammalian cells are the most commonly used production system for therapeutic antibodies. Protocols for the expression of recombinant antibodies in HEK293-6E cells in different antibody formats are described in detail. As model, antibodies against Kallikrein-related peptidase 7 (KLK7) were used. KLK7 is a key player in skin homeostasis and represents an emerging target for pharmacological interventions. Potent inhibitors can not only help to elucidate physiological and pathophysiological functions but also serve as a new archetype for the treatment of inflammatory skin disorders. Phage display-derived affinity-matured human anti-KLK7 antibodies were converted to scFv-Fc, IgG, and Fab formats and transiently prod... More

关键词

Biolayer interferometry, Fab, IgG, Kallikrein-related protease 7, Recombinant antibody formats, Transient gene expression, scFv, scFv-Fc