The optimisation studies used a Central Composite Circumscribed (CCC) design. The DoE studies were of a Central Composite Circumscribed (CCC) design, using a star distance of 1.20 to 1.25 (Fig 3). In this application note only the results of the DoE and verification are described.įig 3. Screening of conditions in PreDictor™ plates (employing a High-throughput Process Development strategy) Table 1 shows the properties of the Dab.ĭuring the set-up of the three-step process, a general workflow was used with the purpose of minimising the development time. With its recombinant protein L ligand, Capto L is a BioProcess™ chromatography medium (resin) with a broad range affinity for antibody fragments of different sizes containing kappa light chains (1).Ĭapto chromatography media were used in a purification process of a kappa subclass Dab expressed in E. The protein L ligand in Capto L binds to the variable region of an antibody’s kappa light chain without interfering with its antigen-binding site.ĭomain antibodies, however, have previously lacked such a platform solution and in this Application note we describe a three-step platform approach using Capto L in the capture step. With the introduction of Capto L (Figure 1), the first industrial platform for the purification of antibody fragments is now emerging. The high purification factor and generic conditions associated with this approach have proven particularly attractive to biopharmaceutical manufacturers.įig 1. The industry standard for purifying MAbs is a platform approach using affinity chromatography with protein A as the capture step. One of the advantages is that due to their structure and smaller size, antibody fragments possess advantageous properties (e.g., easier tissue penetration) that suit a range of diagnostic and therapeutic applications. This platform approach, based on a capture step with Capto L, enables increased efficiency and productivity in developing therapeutics based on Dabs.įollowing on the success of monoclonal antibodies (MAbs), antibody fragments (e.g., Fab, scFv, Dab, etc.) are an increasingly important class of protein-based biotherapeutics. Finally, Capto adhere ImpRes was run in flowthrough mode to render product with undetectable endotoxin levels and ECP content of 80%) over the entire process. Next, a polishing step using Capto MMC ImpRes further reduced ECP and endotoxin levels at yields of > 86%. coli host cell proteins (ECP) and endotoxin levels. First, a capture step using Capto™ L was used to reduce E. This Application note describes a three-step purification process of a domain antibody (Dab) expressed in the periplasm of E.
0 Comments
Leave a Reply. |
AuthorWrite something about yourself. No need to be fancy, just an overview. ArchivesCategories |