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Refolding Technology for scFv Using a New Detergent, N-Lauroyl-L-glutamate and Arginine
Alliance Protein Laboratories, 3957 Corte Cancion, Thousand Oaks, CA 91360, USA
Department of Pharmacology, KEIO University School of Medicine, Tokyo 160-8582, Japan
Institute for Innovation, Ajinomoto Co. Inc., Kanagawa 210-8681, Japan
* Author to whom correspondence should be addressed.
Received: 10 July 2012; in revised form: 16 August 2012 / Accepted: 21 August 2012 / Published: 29 August 2012
Abstract: Monoclonal antibodies to the soluble antigens or cell surface markers hold great promise as effective human therapeutics. One of the major disadvantages is its large size, which prevents efficient penetration into the target tissues. Smaller version of antibodies, which has only antigen binding sites, is extensively investigated. It becomes increasingly apparent, however, that these smaller fragments of antibodies are rather difficult to produce, as the normally efficient mammalian secretion system does not work well for these fragments. Thus, refolding of insoluble proteins produced in Escherichia coli is a method of choice, although such refolding is mainly based on trial-and-error experiment. Here we describe a novel refolding system using a new amino acid-based detergent, N-lauroyl-L-glutamate, and arginine. This detergent appears to readily dissociate from proteins below critical micelle concentration (CMC), while remaining effective in protein solubilization above CMC. Arginine suppresses protein aggregation when the detergent concentration was reduced below CMC. The interaction of the detergent and arginine with proteins, which play an important role in protein refolding, will be discussed in great length.
Keywords: N-lauroyl-L-glutamate; arginine; refolding; scFv; interleukin-6; transglutaminase
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MDPI and ACS Style
Arakawa, T.; Kita, Y.; Ejima, D. Refolding Technology for scFv Using a New Detergent, N-Lauroyl-L-glutamate and Arginine. Antibodies 2012, 1, 215-238.
Arakawa T, Kita Y, Ejima D. Refolding Technology for scFv Using a New Detergent, N-Lauroyl-L-glutamate and Arginine. Antibodies. 2012; 1(2):215-238.
Arakawa, Tsutomu; Kita, Yoshiko; Ejima, Daisuke. 2012. "Refolding Technology for scFv Using a New Detergent, N-Lauroyl-L-glutamate and Arginine." Antibodies 1, no. 2: 215-238.