Author Archives: Judith Vajda

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,…

MAb Aggregate Removal Through Ion-Exchange Chromatography: In Bind/Elute and Flow-Through Mode with TOYOPEARL NH2-750F

A polishing step for the purification of a monoclonal antibody (MAb) using TOYOPEARL NH2-750F was developed. In general, anion-exchange resins can be used in bind and elute (B/E) mode as well as in flow- through (FT) mode. Both options were evaluated. To increase the amount of aggregates of the test sample, a MAb was aggregated by acidic incubation and subsequently diluted to 1 g/L in loading buffer. Read the full text of this application note in the PDF (Login required).

Protein A Chromatography with High-Titer Feedstocks

Protein A chromatography has become a widely used platform in monoclonal antibody (MAb) purification. We describe the impact of higher MAb loadings on aggregation and ligand leaching during protein A chromatography, as well as the benefits of high-capacity Protein A resins for high-titer feedstocks. Today, high MAb titers achieved through advanced upstream processing have become a challenge for downstream processing, especially with regards to the expensive protein A capturing step. TOYOPEARL AF-rProtein A HC-650F provides superior MAb capacity and beneficial…