One of the advantages is that due to their structure and smaller size, antibody fragments possess properties (e.g., easier tissue penetration) that suit a range of diagnostic and therapeutic applications. This platform approach enables increased efficiency and productivity in developing therapeutics based on antibody fragments.Īntibody fragments (e.g., Fab, scFv, domain antibodies, etc.) are set to become the next important class of proteinbased biotherapeutics after monoclonal antibodies (MAbs). Using Capto™ L, Capto SP ImpRes, and Capto Q media in the three step process resulted in efficient removal of the main contaminants and high yields (≥ 87%) over the entire process. This general approach supports Quality by Design (QbD), where the plates give the characterized space and the DoE in columns render both the design and the control space. Design of Experiments (DoE) was used to further optimize the conditions for each step. Next, capacity studies were performed in small columns with purified protein, and elution studies in columns were performed to find the optimal elution pH. First, chromatography media (resins) were screened in 96-well PreDictor™ plates, using wide wash and elution conditions. Published by Elsevier B.V.This Application note describes a three-step purification process of a Fab originating from an E. However, some of this advantage is lost if the feed is a mixture of BSA and Tg since, in this case, Tg binding leads to greater diffusional hindrance for BSA.Ĭore-shell resins Diffusional hindrance Dynamic binding capacity Flow-through purification Modeling.Ĭopyright © 2021. Column measurements show that, despite the higher static capacity of Capto Core 400 for BSA, the dynamic binding capacity is greater for Capto Core 700 as a result of its faster kinetics. Adsorbed Tg further hinders diffusion of BSA in both resins. These values decrease dramatically for Tg to 0.022 × 10 -7 and 0.088 × 10 -7 cm 2/s and to 0.13 × 10 -7 and 0.59 × 10 -7 cm 2/s for Capto Core 400 and 700, respectively. For BSA, core and shell effective pore diffusivities are about 0.25 × 10 -7 and 0.6 × 10 -7 cm 2/s, respectively, for Capto Core 400, and about 1.6 × 10 -7 and 2.6 × 10 -7 cm 2/s, respectively, for Capto Core 700. Mass transfer in both resins is affected by diffusional resistances through the shell and within the adsorbing core. However, for the much larger Tg, the attainable capacity is substantially larger for Capto Core 700. Because of the smaller pores and higher surface area, the BSA binding capacity of Capto Core 400 is approximately double that of Capto Core 700. Although shell thicknesses are comparable (3.6 and 4.2 µm for Capto Core 400 and 700, respectively), the two resins differ substantially in pore size (pore radii of 19 and 50 nm, respectively). Both resins are agarose-based and contain an adsorbing core surrounded by an inert shell. Structural and functional characteristics of the two core-shell resins Capto™ Core 400 and 700, which are useful for the flow-through purification of bioparticles such as viruses, viral vectors, and vaccines, are compared using bovine serum albumin (BSA) and thyroglobulin (Tg) as models for small and large protein contaminants.
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