scFv by Two-Step Cloning

  • Dafne MüllerEmail author
Part of the Springer Protocols Handbooks book series (SPH)


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.


Calf Intestine Alkaline Phosphatase Periplasmatic Expression pelB Leader Antibody Fusion Protein Respective Restriction Enzyme 
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Copyright information

© Springer-Verlag Berlin Heidelberg 2010

Authors and Affiliations

  1. 1.Institute of Cell Biology and ImmunologyUniversity of StuttgartStuttgartGermany

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