Large-scale Chromatography Research Articles

A FAD-SMT Model of Large Scale Liquid Adsorption Chromatography

The paper proposes a FAD-SMT model of large-scale liquid chromatography by which a continuous equation of chromatographic separation is decomposed into a convection dispersion partial differential equation and a set of ordinary differential equations. The numerical method for the FAD-SMT model is established. The stability and the convergence condition of numerical solution, and the choice of time and space interval are discussed. The FAD-SMT model is used to simulate liquid adsorption chromatography and cycling adsorption chromatography. Results show that the elution curves calculated by FAD-SMT model are good agreement with the experimental elution curves of the separation of glucose and fructose, the separation of sucrose and reducing sugar and the separation of mannitol and sorbitol. The result of parameter sensitivity analysis shows that the chromatographic elution curves are more sensitive to the changes of the parameter ai in Langmuir isotherms than to the changes of other parameters in the studied system.

Complement-dependent hemolysis in human serum via the lectin pathway: Regulation by C4bp, factor H and immunoglobulins

A novel method of large-scale chromatography has been developed to improve recovery and purity of immunoglobulin G (IgG) from pooled plasma. The current study compares safety, toxicity and efficacy of two intravenous immunoglobulin products: a novel formulation, IGIV caprylate/chromatography (IGIV-C; Gamunex™, 10%) and a licensed solvent/detergent-treated product, Gamimune®N, 10% (IGIV-SD). The study, a randomized, double-blind, parallel group, therapeutic equivalence trial, was conducted at 25 treatment centers in Canada and the United States. Patients (n=172) having confirmed chronic primary immunodeficiency (PID), aged 1–75 years, and receiving IGIV therapy were enrolled. For 9 months, patients were treated with IGIV-C or IGIV-SD in accordance with the patient's individualized treatment regimen utilized before study entry. The primary endpoint was the proportion of patients with ≥1 validated acute sinopulmonary infection during the treatment period. Secondary endpoints included the proportion of patients with all infections, time to first infection, annual infection rates, lung function parameters, infusion-related safety and viral safety. The annual validated infection rate in the IGIV-C group was 0.18 compared to 0.43 in the IGIV-SD group (p=0.023). Nine patients receiving IGIV-C experienced validated infections, compared to 17 patients in IGIV-SD group (p=0.06). Acute sinusitis (validated plus clinically defined) was less frequent in the IGIV-C group (p=0.012). Presence of bronchiectasis did not affect efficacy. Adverse reactions were similar in frequency and severity in both groups. No evidence of viral transmission was observed. IGIV-C appears to be superior to IGIV-SD in preventing validated sinopulmonary infections, especially acute sinusitis, in patients with PID.

A novel technology for packing and unpacking pilot and production scale columns

A new method of packing and unpacking large scale chromatography columns is described. This involves use of a 3-position valve that can inject chromatography media into a closed column thereby packing it. This same valve in another state is then used to unpack the column. Heights equivalent to a theoretical plate and asymmetry factors of packs on columns from 280 to 800 mm diameter are discussed.

Preparation of lyophilised lipid rich quality control serum by isolating lipids using large scale chromatography technology

The plasma fractionation is a technology to separate therapeutic plasma proteins. The fractionation is carried out either by solvent precipitation (ethanol) or recently by chromatography. During the process of chromatography lipids were found in waste fraction. A new lipid rich lyophilised quality control (Q.C.) sera was prepared inexpensively using this waste plasma fraction. This is probably the first attempt to prepare the Q.C. sera by large scale chromatographic method in India.

High-performance liquid chromatography of amino acids, peptides and proteins CXXXIX. Impact of operating parameters in large-scale chromatography of proteins

Large-scale chromatography has been playing an important role in downstream treatment processing in biotechnology. In order to improve the productivity, the throughput of the chromatographic equipment was often increased by increasing the flow-rate and/or by increasing the column sample loading. This paper reports the results of a study on the impact of these and other operating parameters in affinity and ion-exchange chromatographic columns when used for protein purification. A sectional model was developed to predict protein adsorption processes in a packed column. The formulations of this mathematical model are presented in the Appendix. The present study was carried out with computer simulation based on this model and using data obtained from laboratory-scale columns. This model can simulate both the adsorption and washing stages of the protein purification process for both porous and non-porous particles. The effects of changing operating parameters were simulated and contour plots were generated for the easy identification of these effects. It was shown that both flow-rate and column loading can have a considerable impact on the processing rate and the yield of the column. As for the column capacity utilization, the impact of changing flow-rate is not significant at column loading of less than 80% in the test case. It was suggest that the present investigation provides a systematic predictive strategy which will greatly reduce the need for expensive, labour-intensive and time-consuming experimental work during process scale-up.

High-performance liquid chromatography of amino acids, peptides and proteins CXXXII. Optimisation of operating parameters for protein purification with chromatographic columns

In large-scale chromatography, process optimisation is one of the key elements for success. This paper presents a method for determining the optimum operating parameters for affinity and ion-exchange chromatographic columns when used for protein purification. Based on a mathematical model developed as part of our association investigations, computer programs have been developed to describe the dynamic relationships acting within the chromatographic system. Two basic operating parameters, the flow-rate and the effluent concentration at which the adsorption stage is terminated, can be optimised to give a maximum production rate. The sample loading volume and the processing time then can be determined. The effect of washing conditions on the production rate and the yield is also discussed. Examples are given for a specific system where the optimisation is based on the yield and the percentage utilisation of the column capacity. Contour plots are generated to aid the determination of the range of controlling parameters, and to guide further system design.

Optimization of large-scale chromatography for biotechnological applications

An economic evaluation of a chromatographic separation is discussed. The effects of particle size, cycle time, solvent, and column costs are analyzed. With small particles ( 60 µm), resin costs are less than half of the total. A strong optimum is found between 20–40 µm for maximum productivity, using both Gaussian models and the mass transfer model of Lapidus and Amundson to generate peaks. A new compilation of resin costs, column costs, and pump costs is given.

Modeling of gradient elution in multicomponent nonlinear chromatography

Gradient elution in nonlinear chromatography is a very powerful tool for the separation of components having a wide range of retentivity. It is not only used for chemical analysis, but also used in preparative- and large-scale chromatography for the separation of many macromolecules, such as proteins. In this work, a general rate model has been presented for the study of gradient elution in nonlinear chromatography. The model considers axial dispersion, external film mass transfer, intraparticle diffusion and kinetic effects. The modulator-eluite relationship used in the model accounts for both electrostatic and hydrophobic interactions. Examples of simulations have been given to demonstrate the efficiency and robustness of a computer code based on a numerical procedure for the general rate model, which uses the finite-element, the orthogonal-collocation and Gear's stiff methods. The examples also show the advantage of gradient elution over isocratic elution in multicomponent elutions, and the comparisons among linear, nonlinear and stepwise linear gradients.

Simulation of multicomponent elution with the mobile phase containing competing modifiers

In elution chromatography, modifiers sometimes are added to the mobile phase to compete with sample solutes for binding sites in order to reduce the retention time and band spreading of sample solutes. In this work, the interrelationship between modifiers and sample solutes in preparative and large-scale chromatography is investigated using a nonlinear multicomponent rate model in which axial dispersion, film mass transfer, intraparticle diffusion, and multicomponent Langmuir isotherm are considered. The modifier with a constant concentration in the feed mobile phase is treated as one of the components in the governing equations of the model and is included in the multicomponent Langmuir isotherm. Peaks that are directly attributed to the modifier in the chromatogram are termed system peaks. It has been concluded that for an elution system with two sample solutes and one modifier, there exist three different patterns of system peaks if the sample contains the modifier, and six if the sample does not contain the modifier. It has been found that apart from the thermodynamic effects, the dispersion and mass transfer effects, in some cases, also play important roles in determining system peak patterns. Simulations for an elution system with two modifiers have also been investigated and discussed.

Die Chromatographie, eine zentrale Methode in der biotechnischen Aufarbeitung

AbstractChromatography, an essential method for downstream processing in biotechnology. More than ten years ago, largescale chromatography was established for isolation and purification of high molecular weight natural products, such as proteins and nucleic acids, from fermentation broths and extracts of animal and plant tissues. This was a result of the development of rigid, hydrophilic, and biocompatible polymers with large pore sizes. Compared to the precipitation methods previously used on a large scale, chromatography offers in particular the possibility of producing blood proteins and enzymes of higher purity on a technical scale and without pollution. The various chromatographic methods involve many separation possibilities with excellent selectivities. Progress in improving the chromatographic materials together with advanced equipment were the basis for present‐day importance of chromatography in biotechnology downstream processing.

Large-scale chromatography of recombinant proteins

When recombinant proteins are expressed in bacterial cells and subsequently grown in fermentation tanks, there remains the problem of recovering the product in pure form. The empirical knowledge gained upon recovery of recombinant proteins indicates that a one-step purification process is very unlikely to succeed. However, combinations of modern techniques, such as immunoaffinity chromatography or immobilized-metal affinity chromatography, with classical techniques, such as ion-exchange chromatography, seem to be suitable for large-scale recovery of recombinant proteins.

A review of very large scale chromatography

The relative merits of large scale gas chromatography (GC) and high performance liquid chromatography (HPLC) have not been reviewed for some considerable time. Although methodologies capable of manufacturing high efficiency (and hence cost effective) plant have existed for over a decade, commercial introduction has been remarkably slow. This has primarily been caused by confidentiality restrictions placed on equipment manufacturers, in turn severely limiting wide ranging applications data. These are necessary to raise potential end users confidence that both techniques offer viable large scale routes to separation and purification of mixtures as analysed by laboratory chromatographic methods. Considerable recent progress has been made with large scale HPLC plant, which has led to sufficient published data to make both technical and economic comparisons against GC possible.

The use of the F.p.l.c.R system in method development and process monitoring for industrial protein chromatography

AbstractThe f.p.l.c.R for high performance biomolecular purification is now widely used in laboratory and pilot scale separations, both as a means of developing new methods for purification and for analysing the purity of intermediary and final products. The speed of the technique means that it is possible to analyse fractions from a process step and to receive the results within 15–30 min. Two aspects are examined in this paper: the monitoring of protein product formation during fermentation and the analysis of fractions in large scale chromatography. The suitability of the technique in an operating process is discussed.

Intermediate- and large-scale reversed-phase preparative high-performance liquid chromatography on an axially compressed column: A facile, quantitative separation of 7α- and 7β-methyl-17β-acetoxy-3-oxoandrost-4-enes

Intermediate- and large-scale reversed-phase preparative high-performance liquid chromatography on an axially compressed column: A facile, quantitative separation of 7α- and 7β-methyl-17β-acetoxy-3-oxoandrost-4-enes

A demountable tube for large-scale chromatography and its application to the isolation of hemoglobin A Ic

The construction and operation of a demountable 10 × 100 cm chromatographic tube is described. In a specific application of this apparatus, a load of 50 g of adult human hemoglobin has been chromatographed for the isolation of 2 g of the minor hemoglobin component A Ic.

The coagulative factors of normal human and human cancer tissue

1. Human cancer tissues are characterised by deposition of fibrin. 2. Aqueous extracts of cancer tissue contain labile factors which cause coagulation in buffered diluted human plasma. 3. The coagulative factors of cancer tissue have been fractionated by chromatography on DEAE cellulose into four main groups, A, B, C and D. 4. Comparable extracts of normal human tissues show either negligible coagulative activity or a small amount of activity corresponding roughly to group B. 5. Starch block electrophoresis of cancer tissue extract separates a fast-moving (prealbumin) thermolabile factor chromatographically identical with group C. 6. Extracts of foetal membrane (chorion) contain thermolabile factors corresponding chromatographically with groups B, C and D. 7. The three main coagulative factors of chorion extract have been isolated in active form by large scale chromatography on DEAB cellulose. 8. The nitrogen content of the factors has been found to be significantly lower than in most proteins. 9. The factors have been examined by starch gel electrophoresis. 10. The factors do not coagulate fibrinogen and, therefore, do not contain thrombin.

Effect of Gamma Radiation on Lactose. I. Some Properties of a Reductone Formed in Irradiated Lactose Solution

Summary Formation and characteristics of reductone in gamma-irradiated lactose solution have been studied. Irradiated lactose solution reduced alkaline 2,6-dichlorophenol indophenol and phospho-18-tungstate reagents. The solution also showed a specific absorption spectrum in the ultraviolet region, the maximum wave length of which was shifted to longer wave lengths, and the maximum specific extinction coefficient was increased with decreasing hydrogen ion concentration. A substance detected on paper chromatograms obtained by multiple development using n -butanol, acetic acid, and water (4:1:5) showed the same characteristics as those of the irradiated lactose solution. From the available data on this substance isolated by large-scale paper chromatography, it was tentatively concluded that some reductone-like properties of the irradiated lactose solution can be ascribed to this substance, a kind of reductonate.

Occurrence of beta-aminoisobutyric acid in Mytilus edulis.

beta-Aminoisobutyric acid was isolated from organ extracts of Mytilus edulis. Ion-exchange resins and large-scale paper chromatography were used to isolate minute quantities of the compound. beta-Aminoisobutyric acid was identified by paper chromatography in several solvents and by conversion to DNP-beta-aminoisobutyric acid and subsequent chromatography of the derivative in several solvents.