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Single-step colony assay with autoinduction of scFv expression for the screening of antibody libraries.

BioTechniques. 2019-02; 
KatoMieko, HanyuYos
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Gene Synthesis … for periplasmic protein expression. DNA fragments (Fig. 1A) used for inserting the linker and the restriction sites for cloning V H and V L fragments were synthesized by GenScript (Piscataway, NJ). The generated fragment was … Get A Quote

摘要

We present a simple colony assay method for screening antibody libraries based on autoinduction of antibody fragment expression. This protocol eliminates the need for colony size monitoring and a separate induction step for single-chain Fv (scFv) antibody fragment expression. Here, scFvs are expressed in an automatic and timely fashion during the assay, resulting in high yields of positive clones and substantial time savings. The method was used successfully to establish monoclonal scFvs with high affinity and specificity against human IgG.

关键词

colony assay,monoclonal antibody,scFv,scree