Capturing a scFv from E. coli Using Protein L Affinity Chromatography

Single-chain variable fragments (scFv) are antibodyderived molecules with several advantages over full-length IgGs, including rapid target access and good tissue penetration. Straightforward and efficient capturing solutions similar to protein A for monoclonal antibodies (mAbs) may pave the way for the future success of this class of molecules. Herein we describe general conditions for capturing scFv from Escherichia coli with TOYOPEARL® AF-rProtein L-650F and subsequent optimization to achieve efficient purification.

General Guidelines for CapturinG The κ light chains of mAbs, Fabs, and scFv bind to protein L at neutral pH and physiological conductivity, whereas host-cell proteins (HCPs) flow through. The bound product can then be recovered at pH 2.0–3.5. Generic conditions developed for TOYOPEARL AF-rProtein L-650F are described in Table 1.

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