Experimental characterization of next‐generation expanded‐bed adsorbents for capture of a recombinant protein expressed in high‐cell‐density yeast fermentation

Abstract

Expanded-bed adsorption (EBA) can be particularly useful in protein recovery from high-cell-density fermentation broth where conventional methods for harvest and clarification, such as continuous centrifugation and depth filtration, demand long processing times and are associated with high costs. In this work, the use of next-generation high-particle-density EBA adsorbents, including two mixed-mode resins, for the direct capture of a recombinant protein expressed in yeast at high cell densities is evaluated. Using classical experimental approaches and under different conditions (pH, salt, etc.), Langmuir isotherm parameters for these resins are obtained along with pore diffusivity values. Additional batch adsorption studies with Fastline® MabDirect, the resin that demonstrated the highest static binding capacity for the recombinant protein of interest under the conditions evaluated in this study, indicate competitive binding of nontarget proteins and approximately a 30% reduction in equilibrium binding capacity to 50 mg/mL settled bed in the presence of a 5%-10% cell concentration. Packed-bed (PB) dynamic binding capacities for the MabDirect resin (25-40 mg/mL PB) were significantly higher than for the Fastline® HSA resin and for the MabDirect MM resin in expanded-bed mode (5-10 mg/mL settled bed). Bed expansion behavior for the mMabDirect MM resin along with process yield and eluate purity are identified as a function of linear velocity and cell density, demonstrating process feasibility for pilot scale use.

EFFECTS OF ARTICHOKE (Cynara scolumus L.) EXTRACT ON BIOCHEMICAL PARAMETERS IN DIABETIC RATS

“…The content of total polyphenols found in the artichoke crude extract was similar to the values reported in the literature [Ramaiya, Bujang & Zakaria, 2014]. However, the values found in the artichoke leaves in our study were higher than those which were obtained by [Kelly et al, 2013] (0.83 ± 0.07 g EAG/100g) and less than values found by [Ramaiya, Bujang & Zakaria, 2014] (9.25 g EAG/100g) even in other plant species, such as P. Edulis . This divergence can be attributed to the variations between the different plants, as well as the solvents employed in these studies [Ramaiya, Bujang & Zakaria, 2014].…”

Mathematical modelling of expanded bed adsorption – a perspective on in silico process design

“…A number of authors describe the non‐stationary particle phase by adding an extra equation for the solid phase concentration: where ε s is the solids fraction, q ′ is the average solute concentration in the solid phase and D s is the solids dispersion coefficient. D s is either determined experimentally or is calculated via van der Meer's correlation: …”

Recovery and purification of recombinant 503 antigen of Leishmania infantum chagasi using expanded bed adsorption chromatography

“…Centrifugation the two most widely used techniques for the removal of biomass or cell debris. However, these clarification methods possess several disadvantages, such as being time consuming, resulting in substantial protein loss, and having high operational and capital costs [11,12].…”

“…EBA combines the processes of clarification, concentration and initial purification into a one-step operation and thus provides increased process economy, increased yield, shorter overall processing times, reduced work cost, and reduced running costs and capital expenditure [15][16][17]. EBA has been used successfully with a number of different feedstocks, such as bacterial [18,19], yeast [12,20], and mammalian cell fermentation broths [21,22], as well as others [16,23,24], for the recovery of important biomolecules.…”