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Adsorption of Human Serum Albumin (HSA) on a mixed-mode adsorbent: equilibrium and kinetics

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Abstract

The adsorption equilibrium and kinetics of Human Serum albumin (HSA) onto a novel high particle density multimodal adsorbent were studied by batch adsorption experiments. MabDirect MM, from Upfront Chromatography A/S, is an agarose-tungsten carbide composite adsorbent with an anionic mixed mode ligand. The effects of ionic strength (by addition of salt) and of pH are assessed. Langmuir isotherms parameters are obtained along with pore diffusivity values by fitting the batch experiments using a pore diffusion model. Under the studied conditions for pH 5.0, without salt, it was obtained the largest adsorption capacity (36.0 ± 3.5 mg·gadsorbent,dry −1) a typical behavior for a mixed mode adsorbent. Effective pore diffusivity (Dpe) was estimated as (2.2 ± 0.1) × 10−6 cm2·min−1 for this condition. Additionally, the lowest adsorption capacity (8.6 ± 2.1 mg·gadsorbent,dry −1) and an estimation of Dpe equal to (2.4 ± 0.6) × 10−6 cm2·min−1 were obtained for pH 7.0 without salt. An alternative method of Dpe estimation is used and validate the results obtained by simulations. Furthermore, several experiments were carried out in a fixed bed column with the aim to understand the kinetics and hydrodynamics, and to validate the batch adsorption results. For a feed concentration of ±0.92 g·L−1, pH 5.0, without salt, for cleaned and fresh adsorbent, fixed bed dynamic binding capacities of 13.84 and 14.37 mg·gadsorbent,dry −1 were obtained, respectively, representing near 50% of saturation capacity.

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Abbreviations

Bi:

Biot number, -

\({{C}_{b0}}\) :

Feed protein concentration, g·L−1

\({{C}_{b}}\) :

Protein concentration in the bulk phase, mg·cm−3

\({{C}_{p}}\) :

Protein concentration in the particle pore, mg·cm−3

\({{C}_{NaCl}}\) :

Salt (NaCl) concentration, M

\({{D}_{m}}\) :

Molecular diffusivity, m2·s−1

\({{d}_{p}}\) :

Particle diameter, cm

\({{D}_{pe}}\) :

Effective pore diffusivity, cm2·min−1

\(H\) :

Bed height, cm

\({{k}_{ext}}\) :

External mass transfer coefficient, cm·min−1

\({{k}_{G}}\) :

Global mass transfer coefficient, cm·min−1

\({{k}_{int}}\) :

Internal mass transfer coefficient, cm·min−1

\({{K}_{L}}\) :

Langmuir adsorption constant, L·g−1

\({{M}_{w}}\) :

Molecular weight, g·mol−1

\({{\bar{Q}}_{adsorption}}\) :

Average adsorption flow rate, cm3·min−1

\({{q}_{5\%}}\) :

5% Dynamic binding capacity, mg·cm−3

\(q\) :

Adsorbed concentration, mgprotein·gadsorbent −1

\(\bar{q}\) :

Average adsorbed concentration, mgprotein·gadsorbent −1

\({{q}_{max}}\) :

Maximum adsorption capacity, mgprotein·gadsorbent −1

\(Re\) :

Reynolds number, -

\({{r}_{p}}\) :

Particle radios, cm

\(Sc\) :

Schmidt number, -

\(Sh\) :

Sherwood number, -

\(T\) :

Temperature, K

\({{u}_{i}}\) :

Interstitial velocity, cm·min−1

\(V\) :

Effluent liquid volume from the fixed bed, cm3

\({{V}_{5\%}}\) :

Effluent liquid volume at 5% BSA breakthrough point, cm3

\({{V}_{A}}\) :

Packed bed volume of adsorbent, cm3

\({{V}_{L}}\) :

Volume of protein solution, cm3

\(w\) :

Adsorbent mass, g

\(t\) :

Time, min

\(r\) :

Radial distance, cm

\(z\) :

Axial position, cm

\({{\varepsilon }_{p}}\) :

Particle (solid) porosity, -

\({{\varepsilon }_{b}}\) :

External (bed) porosity, -

\({{\varepsilon }_{T}}\) :

Total particle porosity, -

\(\rho\) :

Density, g·cm−3

\({{\rho }_{\rho }}\) :

Particle density, g·mL−1

\(\tau\) :

Tortuosity, -

\(\eta\) :

Viscosity, cP

HSA:

Human Serum Albumin

BSA:

Bovine Serum Albumin

MM:

Mixed-mode

SDS–Page:

Sodium dodecyl sulfate–Polyacrylamide gel electrophoresis

rpm:

Rotations per minute

pI:

Protein isoelectric point

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Acknowledgements

This work was co-financed by Quadro de Refência Estratégico Nacional (QREN), ON2 and Fundo Europeu de Desenvolvimento Regional (FEDER) (Project NORTE-07-0124-FEDER-000006) and by Fundação para a Ciência e Tecnologia (FCT) and FEDER under Program PT2020 (Project UID/EQU/50020/2013).

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  1. Laboratory of Separation and Reaction Engineering - Laboratory of Catalysis and Materials (LSRE-LCM), Dept. of Chemical Engineering, University of Porto, Rua Dr. Roberto Frias, s/n 4200-465, Porto, Portugal

    Pedro Ferreira Gomes, José Miguel Loureiro & Alírio E. Rodrigues

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  1. Pedro Ferreira Gomes
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  3. Alírio E. Rodrigues
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Correspondence to Alírio E. Rodrigues.

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Gomes, P.F., Loureiro, J.M. & Rodrigues, A.E. Adsorption of Human Serum Albumin (HSA) on a mixed-mode adsorbent: equilibrium and kinetics. Adsorption 23, 491–505 (2017). https://doi.org/10.1007/s10450-017-9861-x

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