scFv by Two-Step Cloning
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The scFv-chain Fv (scFv) corresponds to the smallest functional binding unit of a classical antibody and constitutes a favourite format for the comparative first evaluation of the recombinant potential of an antibody. In this protocol, a two-step cloning strategy is described following the modular exchange principle as a general guideline for converting antibodies into an scFv format suitable for periplasmatic expression in E. coli.
KeywordsCalf Intestine Alkaline Phosphatase Periplasmatic Expression pelB Leader Antibody Fusion Protein Respective Restriction Enzyme
- Gibson TJ (1984) Studies on the Epstein–Barr virus genome. PhD thesis, University of CambridgeGoogle Scholar
- Kim YJ, Neelamegam R, Heo MA, Edwardraja S, Paik HJ, Lee SG (2008) Improving the productivity of single-chain Fv antibody against c-Met by rearranging the order of its variable domains. J Microbiol Biotechnol 6:1186–90Google Scholar
- Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory Press, New YorkGoogle Scholar