SDS-PAGE of soluble and insoluble extracts (5 μl) of the best clones isolated after four rounds of mutagenesis, cloned in the pET23NN plasmid

<p><b>Copyright information:</b></p><p>Taken from "Directed evolution of single-chain Fv for cytoplasmic expression using the β-galactosidase complementation assay results in proteins highly susceptible to protease degradation and aggregation"</p><p>Microbial Cell Factories 2004;3():16-16.</p><p>Published online 17 Dec 2004</p><p>PMCID:PMC544847.</p><p>Copyright © 2004 Philibert and Martineau; licensee BioMed Central Ltd.</p> Proteins were revealed either by Coomassie staining (top) or by western blot using the 9E10 anti-c-myc monoclonal antibody followed by an anti-mouse alkaline phosphatase-conjugated antibody and detected with the chromogenic substrate BCIP/NBT. A) HuLys11 scFv and mutants; B) 225.28S scFv and mutants