Engineering topics

Thormann, Wolfgang

Publications and source records attributed to Thormann, Wolfgang.

11 records

NASA NTRS1987

Experimental and theoretical investigation of the stability of stepwise pH gradients in continuous flow electrophoresis

Isoelectric focusing in the continuous flow mode can be more quickly and economically performed by admitting a stepwise pH gradient composed of simple buffers instead of uniform mixtures of synthetic carrier ampholytes. The time-consuming formation of the pH gradient by the electric field is thereby omitted. The stability of a three-step system with arginine - morpholinoethanesulfonic acid/glycylglycine - aspartic acid is analyzed theoretically by one-dimensional computer simulation as well as experimentally at various flow rates in a continuous flow apparatus. Excellent agreement between experimental and theoretical data was obtained. This metastable configuration was found to be suitable for focusing of proteins under continuous flow conditions. The influence of various combinations of electrolytes and membranes between electrophoresis chamber and electrode compartments is also discussed.

Kuhn, Reinhard↗

NASA NTRS1987

Recent advances in preparative electrophoresis

Various approaches for preparative electrophoresis, and three new instruments for preparative electrophoresis are discussed. Consideration is given to isoelectric focusing, isotachophoresis, and zone electrophoresis, three gel-based electrophoresis methods. The design, functions, and performance of the Elphor VaP 21 device of Hannig (1982), the shear-stabilized BIOSTREAM separator of Thompson (1983), and the recycling isoelectric focusing device are described.

Mosher, Richard A.↗

NASA NTRS1986

The condensation of ampholytes in steady state moving boundaries - Analysis by computer simulation

A digital simulation of the behavior of amphoteric sample components in moving steady state boundaries is presented. Complete computer simulation data, including profiles of concentration, conductivity and pH as functions of time, are given for both cationic and anionic electrolyte configurations which incorporate one amphoteric sample constituent. The condensation of ampholytes in steady state moving boundaries is shown to proceed via an isotachophoretic mechanism and not by isoelectric focusing. Mobility (velocity) relationships necessary for sample components to form steady state zones are discussed.

Mosher, Richard A.↗

NASA NTRS1986

Recent developments in isotachophoresis

Recent advances in the field of isotachophoresis are discussed. These comprise (1) the utilization of digital simulation techniques for computer modeling of isotachophoretic systems of weak electrolytes, (2) the impacts of the models based on migration only, (3) the trend towards automated analytical instruments, and (4) the development of truly industrial-scale isotachophoresis amenable for downstream processing in biotechnology.

Thormann, Wolfgang↗

NASA NTRS1986

An explanation for the plateau phenomenon in isoelectric focusing

Isoelectric focusing of mixtures of simple ampholytes occurs in two phases, an initial rapid separation phase and a second relatively slow stabilizing phase. Transient and steady-state computer simulation data are shown to predict the development of pH plateaus around neutrality during the stabilizing phase of the focusing of such mixtures. This occurs because a nonzero electrophoretic flux is present in a pure zone of focused ampholyte, which is a function of both its isolectric point (pI) and its Delta-pK value. For an ampholyte with a pI greater than 9 or less than 5, this flux causes the development of a significant concentration gradient within its focused zone which is accompanied by a contraction of this zone along the focusing axis. Acidic and basic ampholytes are thereby displayed toward the anode and cathode respectively, creating a pH plateau in the neutral region. Thus, there will be regions of the focusing column, closer to the electrodes and containing more acidic or basic pH values, within which the resolution of samples will reach a maximum and then decrease.

Mosher, Richard A.↗

NASA NTRS1986

Theoretical and experimental separation dynamics in capillary zone electrophoresis

The mathematical model of Bier et al. (1983) is used in a computer aided analysis of the conditions in capillary zone electrophoresis (ZE) under which sample zones migrate noninteractively with the carrier electrolyte. The monitoring of sample zones with a capillary analyzer that features both on-line conductivity and UV detection at the end of the separation trough is discussed. Data from a ZE analysis of a 5-component mixture are presented, and it is noted that all five components can be monitored via their conductivity change if enough sample is present. It is suggested from the results that the concentration ratio of background buffer to sample should be a minimum of 100:1 to effectively apply the plate concept to ZE.

Thormann, Wolfgang↗

NASA NTRS1986

Recycling isotachophoresis - A novel approach to preparative protein fractionation

The concept of automated recycling isotachophoresis (RITP) as a purification methodology is discussed, in addition to a description of the apparatus. In the present automated RITP, the computer system follows the achievement of steady state using arrays of universal and specific sensors, monitors the position of the front edge of the zone structure, activates the counterflow if the leading boundary passes a specified position along the separation axis, or changes the applied current, accordingly. The system demonstrates high resolution, in addition to higher processing rates than are possible in zone electrophoresis or isoelectric focusing.

Sloan, Jeffrey E.↗

NASA NTRS1985

Automation of analytical isotachophoresis

The basic features of automation of analytical isotachophoresis (ITP) are reviewed. Experimental setups consisting of narrow bore tubes which are self-stabilized against thermal convection are considered. Sample detection in free solution is discussed, listing the detector systems presently used or expected to be of potential use in the near future. The combination of a universal detector measuring the evolution of ITP zone structures with detector systems specific to desired components is proposed as a concept of an automated chemical analyzer based on ITP. Possible miniaturization of such an instrument by means of microlithographic techniques is discussed.

Thormann, Wolfgang↗

NASA NTRS1985

The formation of stable pH gradients with weak monovalent buffers for isoelectric focusing in free solution

Two methods which utilize simple buffers for the generation of stable pH gradients (useful for preparative isoelectric focusing) are compared and contrasted. The first employs preformed gradients comprised of two simple buffers in density-stabilized free solution. The second method utilizes neutral membranes to isolate electrolyte reservoirs of constant composition from the separation column. It is shown by computer simulation that steady-state gradients can be formed at any pH range with any number of components in such a system.

Mosher, Richard A.↗

NASA NTRS1985

Electrophoretic transport equations - Electrophoretic models based on migration only and their interrelationships

The general equations which describe the electrophoretic transport of components in solution are restated using Newman's general concept of mobilities. A concise derivation of the moving boundary equation and the regulating function from the continuity equation is presented. Various other regulating principles across moving and stationary boundaries are also discussed, which permits a review of the features and interrelationships of the electrophoretic models based on electromigration only. The effect of considering an interactive (dissociating) solvent on the mathematical treatment is discussed.

Thormann, Wolfgang↗

NASA NTRS1985

A discussion of the constraints which lead to steady state electrophoretic boundaries

The electric field variation across moving boundaries in three component systems is analyzed with a model based on electromigration only (moving boundary equation and regulating function). A decrease in the electric field strength across a boundary in the direction of migration is the most general requirement for the establishment of steady state migrating boundaries. The formation of that electric field gradient is shown to be based on two fundamental constraints. These are (1) the higher net mobility of the leading component compared to that of the terminating constituent and (2) the higher concentration ratio of the leading component to the terminating component ahead of the moving boundary compared to that in the terminating zone. The relations are discussed with respect to strong and weak electrolytes where all three components are present on either side of the boundary. The predictions are verified experimentally. Limitations of the model based on migration only are discussed.

Mosher, Richard A.↗