Arne Staby

Novo Nordisk

The present invention provides a method for industrial-scale protein separation by reverse phase chromatography by use of a buffer system and an additional salt.

The present invention is directed to a method for reducing the onset or incidence of gelation/fibrillation/aggregation during the up- and down-stream processing and purification of peptides, polypeptides and proteins analogs and/or derivatives thereof, including Glucagon-Like Peptides (GLPs). More particularly, the present invention relates to meth...

This report highlights the potential of measurement, monitoring, modeling and control (M(3) C) methodologies in animal and human cell culture technology. In particular, state-of-the-art of M(3) C technologies and their industrial relevance of existing technology are addressed. It is a summary of an expert panel discussion between biotechnologists a...

Small synthetic ligands for protein purification have become increasingly interesting with the growing need for cheap chromatographic materials for protein purification and especially for the purification of monoclonal antibodies (mAbs). Today, Protein A-based chromatographic resins are the most commonly used capture step in mAb down stream process...

The application of high throughput experimentation (HTE) in protein purification process development has created an analytical bottleneck. Using a new label-free and non-invasive methodology for analyzing multicomponent protein mixtures by means of spectral measurements, we show that the analytical throughput for selective protein quantification ca...

Bacterial Protein A (PrtA) and Protein G (PrtG) are widely used for affinity purification of antibodies. An understanding of how PrtA and PrtG bind to different isotypes of immunoglobulin type G (IgG) and to their corresponding Fc fragments is essential for the development of PrtA and PrtG mimetic ligands and for the establishment of generic proces...

Urea is widely employed to facilitate protein separations in ion exchange chromatography at various scales. In this work, five model proteins were used to examine the chromatographic effects of protein conformational changes induced by urea in ion exchange chromatography. Linear gradient experiments were carried out at various urea concentrations a...

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Protein solubility is a key parameter affecting the performance of every biotechnological downstream processing operation. Current methods used to determine protein solubility data, however, are often characterized by the need of pure protein crystals and long experimental time frames lying in the range of weeks to months, while other methods often...

A desired goal of the PAT framework is to design and develop well understood processes that will consistently ensure a predefined quality at the end of the manufacturing process.[ 1 1. Guidance for Industry. PAT–A framework for Innovative Pharmaceutical Development, Manufacturing, and Quality Assurance, US Department of Health and Human Services, F...

This study evaluates the applicability of the liquid handling workstation Tecan Evo Freedom 200 and 200 μL Media Scout RoboColumns to dynamic chromatographic operations such as frontal analysis (breakthrough) and elution experiments in a high throughput screening mode. Breakthrough experiments were conducted using BSA as a model protein and the sta...

A desired goal of the PAT framework is to design and develop well-understood processes that will consistently ensure a predefined quality at the end of the manufacturing process. Such procedures would be consistent with the basic tenet of quality by design and could reduce risks to quality and regulatory concerns while improving efficiency. To supp...

Development of a chromatographic purification step proceeds through a number of stages. High-throughput screening techniques are used to identify suitable resins. This technique is also suitable for the design of a capture step and some intermediate chromatographic steps, but development and true optimisation of a purification step necessitate simu...

A comparative study on weak anion exchangers was performed to investigate the pH dependence, binding strength, particle size distribution, and static and dynamic capacity of the chromatographic resins. The resins tested included: DEAE Sepharose FF, Poros 50 D, Fractogel EMD DEAE (M), MacroPrep DEAE Support, DEAE Ceramic HyperD 20, and Toyopearl DEA...

Cross-flow membrane microfiltration was used under optimal conditions to recover met-growth hormone inclusion bodies (IBs) from Escherichia coli cell lysate by removal of the host-cell (bacterial) proteins (HCP) under minimal fouling conditions. This is the first step of a two-step process in which the goal was to isolate IBs at high yield from the...

Strong and weak cation-exchangers were compared for a number of chromatographic parameters, i.e. pH dependence, efficiency, binding strength, particle size distribution, static and dynamic capacity, and scanning electron microscopy (SEM) pictures. Chromatographic resins investigated were Fractogel EMD SO3- (M), Fractogel EMD SE Hicap (M), Fractogel...

A comparative study was performed on heparin resins and strong and weak cation exchangers to investigate the pH dependence, efficiency, binding strength, particle size distribution, static and dynamic capacity, and scanning electron microscopy pictures of chromatographic resins. The resins tested include: Heparin Sepharose FF, SP Sepharose FF, CM S...

A comparative study was performed on strong cation-exchangers to investigate the pH dependence, efficiency, binding strength, particle size distribution, static and dynamic capacity, and SEM pictures of chromatographic resins. The resins tested included: SP Sepharose XL, Poros 50 HS, Toyopearl SP 550c, SP Sepharose BB, Source 30S, TSKGel SP-5PW-HR2...

A comparative study was performed on strong anion exchangers to investigate the pH dependence, titration curves, efficiency, binding strength, particle size distribution, and static and dynamic capacity of the chromatographic resins. The resins tested included Q Sepharose XL, UNO Q-1, Poros 50 HQ, Toyopearl QAE 550c, Separon HemaBio 1000Q, Q-Cellth...

A comparative study has been undertaken on various strong anion-exchangers to investigate the pH dependence, titration curves, efficiency, binding strength, and dynamic capacity of the chromatographic resins. The resins tested included: Macro-Prep 25Q, TSK-Gel Q-5PW-HR, Poros QE/M, Q Sepharose FF, Q HyperD 20, Q Zirconia, Source 30Q, Fractogel EMD...

Chromatographic media suppliers most frequently state the capacities of their gels based on either static capacities or frontal analysis experiments of pure proteins, however, these capacity values are often far from the capacities experienced in the production of such proteins. In this work static and dynamic capacities of various pure industrial...

Perfusion chromatography is a rather new HPLC concept using packing materials having large through-pores, which were developed to overcome the problems of stagnant mobile mass transfer without loss of separation speed and column capacity. The perfusion media have been developed for both analytical and preparative separations in a number of distinct...

Phase equilibria of 1-butanol in carbon dioxide, ethene, ethane, and propane have been investigated at a reduced temperature of 1.03 of the supercritical gases corresponding to 290.8 K for ethene, 313.4 K for carbon dioxide, 314.4 K for ethane, and 381.0 K for propane and pressures from 1.60 to 8.31 MPa. Experimental and smoothed phase compositions...

Measurements of phase equilibria of urea-fractionated fish oil fatty acid ethyl esters (FAEEs) in supercritical carbon dioxide were performed at pressures from 8 to 26 MPa and temperatures of 313.2 and 343.2 K. Experimental temperatures, pressures, and mass fractions are reported in tabular form. The equilibrium ratios of the esters depend strongly...

Various natural and processed fish oil triglyceride mixtures have been analyzed by capillary supercritical fluid chromatography (SFC). The analyses were performed on nonpolar columns to separate the components by lipid class and by the number of carbon atoms. The compounds separated included free fatty acids, squalene, α-tocopherol, cholesterol, wa...

Supercritical fluid chromatographic analysis methods have been employed in the examination of several marine oils for the group separation of free fatty acids, retinol, ergocalciferol, cholecalciferol, squalene, tocopherols, cholesterol, was esters, diacylglycerols, cholesteryl esters, and triacylglycerols. The oils were derived from characteristic...

Solubilities and K-values of urea-fractionated fish oil fatty acid ethyl esters, prepared from sandeel oil, have been measured in supercritical carbon dioxide experimentally. The measurements were performed at the temperatures of 313.2 and 343.2 K in the pressure range of 8 to 26 MPa. Experimental temperatures, pressures, and densities of the equil...

The use of supercritical fluids, especially carbon dioxide, in separation of fish oil and derived compounds has increased the last decade due to the expected advantages of supercritical separation processes. In pursuit of the design of supercritical fluid separation processes, phase equilibria and pilot plant data are essential. In this paper we pr...

A fish oil fatty acid ethyl ester mixture derived from the sand eel (Ammodytes lancea) has been analysed using various capillary gas chromatography (GC) and supercritical fluid chromatography (SFC) methods and 39 components including cholesterol have been identified. The results of the analytical SFC and GC experiments are compared showing a good r...

Measurements of phase equilibria of fish oil fatty acid ethyl esters in sub- and supercritical carbon dioxide were performed at pressures from 2 to 22 MPa at the temperatures 283.2, 313.2, and 343.2 K. Experimental temperatures, pressures, mass fractions, mole fractions, equilibrium ratios, and densities are reported. The K-values of the esters dep...

A list of references from a literature retrieval presenting experimental mutual solubilities and phase equilibria of binary systems of carbon dioxide and monoalcohols is given. The list includes the pressure and temperature ranges, the number of data points, and the references. The literature retrieval includes references to the middle of 1992.

The isothermal phase equilibria of 1-pentanol/carbon dioxide have been measured in a variable volume static cell-type apparatus, at pressures from 2 to 18 MPa at the temperatures of 283.2, 313.2, 343.2, and 373.2 K. Experimental temperatures, pressures, mole fractions, and densities are reported.

Thesis (doctoral)--University of Denmark, 1993.

An apparatus for measurement of solubilities of liquids in supercritical fluids has been constructed. It consists of a 70-MPa (10,000-psia) PVT cell where supply systems for liquids and gases have been added. The cell is a pistoned 130-cc pressure-compensated glass tube, where stirring is accomplished by rocking the cell. A liquid and a supercritic...

The volumetric properties of a nitrogen-methane-ethane mixture are determined experimentally by a Burnett apparatus at 275, 310, and 345 K at pressures to 60 MPa. In this paper compression factors are calculated by fitting simultaneously the isothermal pressure series to the Burnett relation and the virial equation. Densities and compression factor...