Continuous aqueous two-phase extraction of human antibodies using a packed column

The performance of a pilot scale packed differential contactor was evaluated for the continuous counter-current aqueous two-phase extraction (ATPE) of human immunoglobulin G (IgG) from a Chinese hamster ovary (CHO) cells supernatant (CS) enriched with pure protein. Preliminary studies have been firstly performed in order to select the dispersed phase (phosphate-rich or polyethylene glycol 3350 Da (PEG)-rich phase) and the column packing material. The PEG-rich phase has been selected as the dispersed phase and the stainless steel as the preferred material for the column packing bed since it was not wetted preferentially by the selected dispersed phase. Hydrodynamic studies have been also performed, and the experimental results were successfully adjusted to the Richardson-Zaki and Mísek equations, typically used for the conventional organic-aqueous two-phase systems. An experimental set-up combining the packed column with a pump mixer-settler stage showed to have the best performance and to be advantageous when compared to the IgG batch extraction. An IgG recovery yield of 85% could be obtained with about 50% of total contaminants and more than 85% of contaminant proteins removal. Mass transfer studies have revealed that the mass transfer was controlled by the PEG-rich phase. A higher efficiency could be obtained when using an extra pump mixer-settler stage and higher flow rates.     

Cadmium removal in a biosorption column

New biosorbent material derived from a ubiquitous brown marine alga Ascophyllum nodosum has been examined in packed-bed flow-through sorption columns. It effectively removed 10 mg/L of cadmium down to 1.5 ppb levels in the effluent, representing 99.985% removal. The experimental methodology used was based on the early Bohart and Adams sorption model, resulting in quantitative determination of the characteristic process parameters which can be used for performance comparison and process design. An average metal loading of the biosorbent (N(0)) determined was 30 mg Cd/g, corresponding closely to that observed for the batch equilibrium metal concentration of 10 mg Cd/L. The critical bed depth (D(min)) for the potable water effluent quality standard (0.005 mgg Cd/L) varied with the column feed flow rate (2.4 to 9.6 L/h . cm(2)) from 20 to 50 cm. The sorption column mass transfer and dispersion coefficients were determined, which are also required for solving the sorption model equations. (c) 1994 John Wiley & Sons, Inc.     

Continuous countercurrent salting-out extraction of 1,3-propanediol from fermentation broth in a packed column

The possibility of continuous extraction of 1,3-propanediol in a experimental packed column was investigated using a salting-out extraction system of dipotassium phosphate/ethanol. Mass transfer of 1,3-propanediol takes place from the dispersed phase (salt-rich solution) to the continuous phase (ethanol). The influences of flow rate of dispersed phase and size of packing material on partition coefficient and recovery of 1,3-propanediol were investigated and the results were compared with those obtained in spray column and test tube. Furthermore, the influences of various system compositions on hold up of dispersed phase, mass transfer coefficient, and system stability were also studied in the column packed by stainless steel Dixon 3 × 3 mm. It was found that the packed column showed a good extraction efficiency and stability. Besides, 1,3-propanediol recovery of 90.30% was obtained during a 11 h continuous operation when the real fermentation broth was used. At the same time, 94.4% of phosphate could be recovered when 0.2 volume of anhydrous ethanol was added into the raffinate phase at pH 4.0.     

Continuous removal of malathion by immobilised biomass of Bacillus species S14 using a packed bed column reactor

Abstract

Biosorption of malathion from aqueous solution was studied using Bacillus sp. S₁₄ immobilised on calcium alginate (3%) using a packed bed column reactor at a temperature of 25 °C and a pH of 7.0. The experiments were conducted to study the effect of important design parameters such as bed height, flow rate and influent malathion concentration. Maximum removal capacity (57%) was found at 4 mL min^-1 flow rate, 6.0 cm bed height and 25 mg L^-1 influent malathion concentration. The Adam-Bohart model, Wolborska model, Thomas model, Yoon-Nelson model were employed to determine characteristic parameters such as saturation concentration, external mass transfer coefficient, Thomas rate constant, the maximum solid phase concentration of the solute, rate constant, and the time required for 50% adsorbate breakthrough time, which are all useful for process design. Experimental data were well fitted with Adam–Bohart model at the lower region of effluent/influent malathion concentration values but at higher region values data fitted well with the Thomas and Yoon-Nelson models.     

Heavy metal removal in a biosorption column by immobilized M. rouxii biomass

Mucor rouxii biomass was immobilized in a polysulfone matrix. The spherical immobilized biomass beads were packed in a column. The biosorption column was able to remove metal ions such as Pb, Cd, Ni and Zn not only from single-component metal solutions but also from multi-component metal solutions. Column kinetics for metal removal were described by the Thomas model. For single-component metal solutions, the metal removal capacities of the beads for Pb, Cd, Ni and Zn were 4.06, 3.76, 0.36 and 1.36 mg/g, respectively. For a multi-component metal solution containing Cd, Ni and Zn, the capacities were 0.36, 0.31 and 0.40 mg/g for Cd, Ni and Zn, respectively. The adsorbed metal ions were easily desorbed from the beads with 0.05N HNO3 solution. After acid desorption and regeneration with deionized water, the beads could be reused to adsorb metal ions at a comparable capacity.     

Alkaloid formation by Catharanthus roseus cells in a packed column biofilm reactor

Summary Catharanthus roseus cells producing indole alkaloids were grown on surfaces of Ca-alginate beads within the interspacial volume of a packed column. Production media was circulated through the packed column in an upflow mode. Growth and indole alkaloid formation were quantified and compared with suspension culture of cells. Final alkaloid concentration and alkaloid yield obtained in the packed bed was superior to those obtained in suspension culture. This is thought to be due to improved cell-cell contact and interaction in the packed column.     

Removal of cadmium ions in wastewater through biosorption

Cadmium ions in wastewater can be removed by biosorption on to Acinetobacter calca var. antratus, isolated from a wastewater sample from a copper refinery. There are two equillibrium states: the first, which is reached quickly, is probably direct adsorption, and the second may be due to the release of ion adsorption materials by lysed cells.The authors are with the Department of Chemical Engineering, National Taiwan University, Taipei, Taiwan 10764, ROC.     

Process development of continuous hydrogen production by Enterobacter aerogenes in a packed column reactor

Hydrogen bioproduction from agro-industrial residues by Enterobacter aerogenes in a continuous packed column has been investigated and a complete reactor characterization is presented. Experimental runs carried out at different residence time, liable of interest for industrial application, showed hydrogen yields ranging from 1.36 to 3.02 mmol_H2mmol^у_glucose or, in other words, from 37.5% to 75% of the theoretical hydrogen yield. A simple kinetic model of cell growth, validated by experimental results and allowing the prediction of biomass concentration profile along the reactor and the optimization of superficial velocity, is suggested. By applying the developed approach to the selected operative conditions, the identification of the optimum superficial velocity v_0,opt of about 2.2 cm h^у corresponding to the maximum hydrogen evolution rate H£_2g,max, was performed.     

Batch and column studies of biosorption of heavy metals by Caulerpa lentillifera

The biosorption of Cu(II), Cd(II), and Pb(II) by a dried green macroalga Caulerpa lentillifera was investigated. The sorption kinetic data could be fitted to the pseudo second order kinetic model. The governing transport mechanisms in the sorption process were both external mass transfer and intra-particle diffusion. Isotherm data followed the Sips isotherm model with the exponent of approximately unity suggesting that these biosorption could be described reasonably well with the Langmuir isotherm. The maximum sorption capacities of the various metal components on C. lentillifera biomass could be prioritized in order from high to low as: Pb(II)>Cu(II)>Cd(II). The sorption energies obtained from the Dubinin-Radushkevich model for all sorption systems were in the range of 4-6 kJ mol(-1) indicating that a physical electrostatic force was potentially involved in the sorption process. Thomas model could well describe the breakthrough data from column experiments. Ca(II), Mg(II), and Mn(II) were the major ions released from the algal biomass during the sorption which revealed that ion exchange was one of the main sorption mechanisms.     

Kinetics of biosorption of cadmium on Baker's yeast

In the present study the kinetics of biosorption of cadmium(II) ions by deactivated protonated yeast converted to sodium form was investigated for different initial concentrations of the metal ion (10-100 ppm) and different sorbent dosages (0.1-2.0 g) at a pH of 6.5. The adsorption process occurred in four distinct steps and the rates for these steps decreased sequentially. The rate of cadmium uptake in each case was pseudo-second-order with respect to metal ion concentration. The amount sorbed at equilibrium was found to be directly proportional to the initial metal ion concentration divided by the sorbent mass.     

Repeated cadmium biosorption by regeneratedEnterobacter aerogenes biofilm attached to activated carbon

Summary The bacteriumEnterobacter aerogenes has been used to develop a biofilm over activated carbon for biosorption from various strength cadmium solutions (25–500ppm). High bacterial resistance to metal poisoning allowed biofilm regeneration to raise the net loading of cadmium over the carbon by repeated biosorption runs.     

Comparison of cadmium biosorption by Gram-positive and Gram-negative bacteria from activated sludge

Summary Gram-positive and Gram-negative bacteria were isolated from activated sludge and used to evaluate differences in cadmium biosorption. Gram-positive bacteria exhibited approximately 20% more cadmium biosorption at 30°C and pH 6.6 than Gram-negatives. Biosorption was largely passive in both cases, although metabolic uptake appeared to occur to a higher extent with Gram-positive bacteria.     

Optimization of solid-state fermentation of citrus dried peel by aspergillus niger in a packed bed column

The protein enrichment of citrus dried peel on solid-state fermentation by Aspergillus niger in packed bed column was optimized using a two factor central composit design with α = 2 considering the “initial moisture content of the fermentation mass” and the “temperature in the centre of the fermenter”. It results significance for the moisture content (optimum 56%) and non significance for the temperature in the conditions employed. This indicates that a considerable range of temperature increase for the investigated SSF system could be allowed.     

Direct enantiomeric resolution of ibuprofen and flurbiprofen by packed column SFC

Packed column SFC is used to resolve the enantiomers of two nonsteroidal antiinflammatories. A Chiralpak AD column is used with a binary mobile phase of carbon dioxide/methanol to achieve resolution. The effects of composition, pressure, temperature, and flow on ibuprofen enantiomer separation are examined. In this instance, temperature affords the greatest change in resolution followed by pressure and composition. Baseline resolution of ibuprofen is achieved in less than 7 min and fluibiprofen is baseline resolved in less than 4 min. © 1994 Wiley-Liss, Inc.     

Biosorption of copper by marine algae Gelidium and algal composite material in a packed bed column

Marine algae Gelidium and algal composite material were investigated for the continuous removal of Cu(II) from aqueous solution in a packed bed column. The biosorption behaviour was studied during one sorption–desorption cycle of Cu(II) in the flow through column fed with 50 and 25 mg l⁻¹ of Cu(II) in aqueous solution, at pH 5.3, leading to a maximum uptake capacity of ≈13 and 3 mg g⁻¹, respectively, for algae Gelidium and composite material. The breakthrough time decreases as the inlet copper concentration increases, for the same flow rate. The pH of the effluent decreases over the breakthrough time of copper ions, which indicates that ion exchange is one of the mechanisms involved in the biosorption process. Temperature has little influence on the metal uptake capacity and the increase of the ionic strength reduces the sorption capacity, decreasing the breakthrough time. Desorption using 0.1 M HNO₃ solution was 100% effective. After two consecutive sorption–desorption cycles no changes in the uptake capacity of the composite material were observed. A mass transfer model including film and intraparticle resistances, and the equilibrium relationship, for adsorption and desorption, was successfully applied for the simulation of the biosorption column performance.     

Mass transfer in aqueous two-phases system packed column

SCH 201781 is a direct thrombin inhibitor recently under study in clinical trials to determine its safety and efficacy for the treatment of venous and arterial thrombosis. In aqueous solution, SCH 201781 exists as three forms, a ring-opened hydrated form and two ring-closed diastereomers. An automated solid-phase extraction LC-MS/MS method that chromatographically separates and measures each form was developed and validated from 1 to 1000 ng/mL in human plasma. For calibration curve standards, within- and between-run precision (%CV) ranged from 0.6 to 13.7%, while accuracy (%bias) ranged from -4.8 to 13.1%. For quality control samples, within- and between-run %CV ranged from 1.5 to 9.9% while %bias ranged from -9.1 to 4.9%. The method requires a sample volume of 0.8 ml and utilizes 2H6-labeled SCH 201781 as the internal standard. For sample processing, an Isolute C-8 96-well solid phase extraction plate and a Tomtec Quadra 96 sample processor is employed. Separation of the three forms of SCH 201781 is achieved using a 5 microm, 2 mm x 100 mm Asahipak C8 HPLC column and gradient elution. A Sciex API 365 equipped with a turbo ionspray source is used in the selected reaction monitoring mode for detection. The validated method was used to support clinical studies.     

Estimation of mass transport parameters in a column packed with BioSep beads

An approach to estimate the parameters needed to describe the performance of a column packed with BioSepTM beads is presented. These parameters, which characterize the transport of materials through the column, have been estimated from experimental residence time distribution (RTD) data using both porous and non-porous beads. A laboratory-scale column was used to obtain the experimental data using several ionic species. © Rapid Science Ltd. 1998     

Interparticle mass transfer study with a packed column of immobilized microbes

The kinetics of whole-cell lactase of Escherichia coli immobilized in spherical agar gels was determined under the influence of interparticle mass transfer in a fixed bed reactor. The dependence of the pseudokinetic parameters (K′_m and v′_max) fluid conditions was in accordance with the prediction derived from combinedeffects of kinetics and mass transfer. Within the limited conditions of theexperiments, it was observed that the apparent Michaelis constant could be represented by the following simple equation, K′_m/K_m = 1 + (?/k_La)(v_max/K_m). The mass transfer coeflicient (k_L) needed in the theoretical analysis was determined in correlation with particle size (0.12–0.45 cm) and liquid flow rate (0.30–12.0 cm/min) using the more stable enzyme, cell-bound invertase of Saccharomyces pastorianus, as the material to be entrapped in the gelatinous particles. The relationship was expressed in the following dimensionless equation, (1–?)Sh = 2 + 0.6Re^1/2Sc^1/3, except that marked deviation resulting in the reduction of k_L was noticed with a Reynolds number less than 0.8.     

Aroma compounds produced by Kluyveromyces marxianus in solid state fermentation on a packed bed column bioreactor

Studies were carried out for the production of aroma compounds by Kluyveromyces marxianus grown on cassava bagasse in solid state fermentation using packed bed reactors, testing two different aeration rates. Respirometric analysis was used to follow the growth of the culture. Headspace analysis of the culture by gas chromatography showed the production of 11 compounds, out of which nine were identified. Ethyl acetate, ethanol and acetaldehyde were the major compounds produced. Lower aeration rate (0.06l h^–1 g^–1 of initial dry matter) increased total volatile (TV) production and the rate of production was also increased at this aeration rate. Using an aeration rate of 0.06l h^–1 g^–1 maximum TV concentrations were reached at 24 h and at 40 h with 0.12l h^–1 g^–1.