Engineering topics

Snyder, Robert S.

Publications and source records attributed to Snyder, Robert S..

At least 37 records · Page 2

NASA NTRS1989

Compact Apparatus Grows Protein Crystals

Laboratory apparatus provides delicately balanced combination of materials and chemical conditions for growth of protein crystals. Apparatus and technique for growth based on hanging-drop method for crystallization of macromolecules. Includes pair of syringes with ganged plungers. One syringe contains protein solution; other contains precipitating-agent solution. Syringes intrude into cavity lined with porous reservoir material saturated with 1 mL or more of similar precipitating-agent solution. Prior to activation, ends of syringes plugged to prevent transport of water vapor among three solutions.

Bugg, Charles E.↗

NASA NTRS1989

Electrohydrodynamic distortion of sample streams in continuous flow electrophoresis

Continuous flow electrophoresis experiments were carried out, using an electrolyte and a sample both made of aqueous solutions of phosphate buffer (with polystyrene latex added for visibility), to investigate causes of the sample spreading in this procedure. It is shown theoretically that an electric field perpendicular to a circular filament of conducting fluid surrounded by a fluid of different conductivity produces an electrohydrodynamic flow, which distorts the filament into an ellipse. Experimental results were found to be fully consistent with theretical predictions. It was found that the rate of distortion of the sample stream into a ribbon was proportional to the square of the applied voltage gradient. Furthermore, the orientation of the ribbon depends on the ratios of dielectric constant and electrical conductivity between the buffer and the sample.

Rhodes, Percy H.↗

NASA NTRS1989

Reduced gravity - A new biomedical research environment

Experiment programs for continuous flow electrophoresis and protein crystal growth are described to demonstrate the utility of the reduced gravity environment for scientific research. The advantages of the reduced gravity environment are outlined. The results of experiments on the Space Shuttle using the Continuous Flow Electrophoresis System and crystal growth experiments on Spacelab-1 and the Space Shuttle are examined, noting the importance of microgravity research.

Snyder, Robert S.↗

NASA NTRS1989

Application of partition technology to particle electrophoresis

The effects of polymer-ligand concentration on particle electrophoretic mobility and partition in aqueous polymer two-phase systems are investigated. Polymer coating chemistry and affinity ligand synthesis, purification, and analysis are conducted. It is observed that poly (ethylene glycol)-ligands are effective for controlling particle electrophoretic mobility.

Van Alstine, James M.↗

NASA NTRS1989

Cell separation in immunoaffinity partition in aqueous polymer two-phase systems

Two methods for immunoaffinity partitioning are described. One technique involves the covalent coupling of poly (ethylene glycol) (PEG) to immunoglobulin G antibody preparations. In the second method PEG-modified Protein A is used to complex with cells and unmodified antibody. The effects of PEG molecular weight, the degree of modification, and varying phase system composition on antibody activity and its affinity for the upper phase are studied. It is observed that both methods resulted in effective cell separation.

Karr, Laurel J.↗

NASA NTRS1988

Continuous flow electrophoresis system experiments on shuttle flights STS-6 and STS-7

The development of a space continuous flow electrophoresis system (CFES) is discussed. The objectives of the experiment were: (1) to use a model sample material at a high concentration to evaluate the continuous flow electrophoresis process in the McDonnell Douglass CFES instrument and compare its separation resolution and sample throughput with related devices on Earth, and (2) to expand the basic knowledge of the limitations imposed by fluid flows and particle concentration effects on the electrophoresis process by careful design and evaluation of the space experiment. Hemoglobin and polysaccharide were selected as samples of concentration effects. The results from space show a large band spread of the high concentration of the single species of hemoglobin that was principally due to the mismatch of electrical conductivity between the sample and buffer.

Snyder, Robert S.↗

NASA NTRS1988

Preparative electrophoresis for space

A premise of continuous flow electrophoresis is that removal of buoyance-induced thermal convection caused by axial and lateral temperature gradients results in ideal performance of these instruments in space. Although these gravity dependent phenomena disturb the rectilinear flow in the separation chamber when high voltage gradients or thick chamber are used, distortion of the injected sample stream due to electrodynamic effects cause major broadening of the separated bands. The electrophoresis separation process is simple, however flow local to the sample filament produced by the applied electric field were not considered. These electrohydrodynamic flows distort the sample stream and limit the separation. Also, electroosmosis and viscous flow combine to further distort the process. A moving wall concept is being proposed for space which will eliminate and control the disturbances. The moving wall entrains the fluid to move as a rigid body and produces a constant residence time for all samples distributed across the chamber thickness. The moving wall electrophoresis chamber can only be operated in space because there is no viscous flow in the chamber to stabilize against thermal convection.

Rhodes, Percy H.↗

NASA NTRS1988

Preliminary observations of the effect of solutal convection on crystal morphology

Studies to examine the effect of solutal convection on crystal morphology using sucrose as a model system were initiated. Aspect ratios, defined as the width of the 100-plane-oriented face over the width of the 001-plane-oriented face, were determined for oriented crystals which were grown with either the 001-oriented or the 100-oriented face perpendicular to the convective flow. The dependence of the crystal morphology on orientation is much greater for crystals grown with one face occluded than for crystals grown suspended in solution. Many factors appear to interact in a complex fashion to influence crystal morphology.

Broom, M. Beth H.↗

NASA NTRS1988

Moving wall, continuous flow electronphoresis apparatus

This invention relates generally to electrophoresis devices and more particularly to a moving wall, continuous flow device in which an electrophoresis chamber is angularly positionable with respect to the direction of moving belt walls. A frame with an electrophoresis chamber is rotatably supported between two synchronously driven belt walls. This allows the chamber to be angularly positionable with respect to the direction of belt travel, which compensates for electroosmotic flow within the electrophoresis chamber. Injection of a buffer solution via an opening and a homogenous sample stream via another opening is performed at the end of a chamber, and collection of buffer and the fractionated species particles is done by a conventional collection array at an opposite end of the chamber. Belts are driven at a rate which exactly matches the flow of buffer and sample through the chamber, which entrains the buffer to behave as a rigid electrophoretic medium, eliminating flow distortions (Poiseuille effect). Additionally, belt material for each belt is stored at one end of the device and is taken up by drive wheels at an opposite end. The novelty of this invention particularly lies in the electrophoresis chamber being angularly positionable between two moving belt walls in order to compensate for electroosmotic flow. Additionally, new belt material is continuously exposed within the chamber, minimizing flow distortion due to contamination of the belt material by the sample.

Rhodes, Percy H.↗

NASA NTRS1988

Non-contact temperature measurements for biotechnology discipline working group

In the biotechnology research areas, there is interest in measuring temperature changes over very small dimensions, such as the surface of a 10-micrometer diameter biological cell immersed in cell culture fluid. Non-interference measurements of other properties, such as chemical constituents and their concentrations, are also needed. Contacting probes for pH have recently been developed to penetrate a cell but questions have been raised about their accuracy and net value.

Snyder, Robert S.↗

NASA NTRS1988

Cell separation by immunoaffinity partitioning with polyethylene glycol-modified Protein A in aqueous polymer two-phase systems

Previous work has shown that polyethylene glycol (PEG)-bound antibodies can be used as affinity ligands in PEG-dextran two-phase systems to provide selective partitioning of cells to the PEG-rich phase. In the present work it is shown that immunoaffinity partitioning can be simplified by use of PEG-modified Protein A which complexes with unmodified antibody and cells and shifts their partitioning into the PEG-rich phase, thus eliminating the need to prepare a PEG-modified antibody for each cell type. In addition, the paper provides a more rigorous test of the original technique with PEG-bound antibodies by showing that it is effective at shifting the partitioning of either cell type of a mixture of two cell populations.

Karr, Laurel J.↗

NASA NTRS1988

Thermally reversible gels in electrophoresis. I - Matrix characterization

Two series of thermally reversible hydrogen-bonded gels have been characterized: (5 pct) PVA-(4 pct) PEG and (5 pct) PVA-(0.04 pct) borate gels. They both have extremely low melting points (16-17 C) and could be of potential interest for recovery of proteins after preparative electrophoresis. The PVA-borate gels can be exploited in the pH range 7-11 by progressively increasing the borate content in the pH interval 8 to 7 and concomitantly decreasing the borate levels in the pH zone 8 to 11. It is hypothesized that the low melting point of these gels is due to the fact that they are sparingly and sparsely hydrogen bonded along the PVA chain: on the average, 1 OH group out of 3 or 4 OH groups in the PVA polymer should be engaged in H-bond formation.

Righetti, Pier Giorgio↗

NASA NTRS1987

Preparative electrophoresis for space

A premise of continuous flow electrophoresis is that removal of buoyancy-induced thermal convection caused by axial and lateral temperature gradients results in ideal performance of these instruments in space. Although these gravity dependent phenomena disturb the rectilinear flow in the separation chamber when high voltage gradients or thick chambers are used, distortion of the injected sample stream due to electrohydrodynamic effects cause major broadening of the separated bands. The electrophoresis separation process is simple, however flow local to the sample filament produced by the applied electric field have not been considered. These electrohydrodynamic flows distort the sample stream and limit the separation. Also, electroosmosis and viscous flow combine to further distort the process. A moving wall concept is being proposed for space which will eliminate and control the disturbances. The moving wall entrains the fluid to move as a rigid body and produces a constant residence time for all samples distributed across the chamber thickness. The moving wall electrophoresis chamber can only be operated in space because there is no viscous flow in the chamber to stabilize against thermal convection.

Rhodes, Percy H.↗

NASA NTRS1987

Continuous flow electrophoresis system experiments on shuttle flights STS-6 and STS-7

A space continuous flow electrophoresis system (CFES) was developed that would incorporate specific modifications to laboratory instruments to take advantage of weightlessness. The specific objectives were to use a model sample material at a high concentration to evaluate the continuous flow electrophoresis process in the CFES instrument and compare its separation resolution and sample throughput with related devices on Earth and to expand the basic knowledge of the limitations imposed by fluid flows and particle concentration effects on the electrophoresis process by careful design and evaluation of the space experiment. Hemoglobin and polysaccharide were selected as primary samples. The results from space show a large band spread of the high concentration of the single species of hemoglobin that was due to the mismatch of electrical conductivity between the sample and the buffer. On STS-7 the major objective was to evaluate the influence of the electrical properties of the sample constituents on the resolution of the CFES. As expected, the polystyrene latex microspheres dispersed in a solution with 3 times the electrical conductivity of the curtain buffer separated with a larger band spread than in the 2nd experiment.

Snyder, Robert S.↗

NASA NTRS1987

Compensating for Electro-Osmosis in Electrophoresis

Simple mechanical adjustment eliminates transverse velocity component. New apparatus for moving-wall electrophoresis increases degree of collimation of chemical species in sample stream. Electrophoresis chamber set at slight angle in horizontal plane to adjust angle between solution flow and wall motion. Component of velocity created cancels electro-osmotic effect.

Rhodes, Percy H.↗

NASA NTRS1987

Liquid drop stability for protein crystal growth in microgravity

It is possible to grow protein crystals for biomedical research in microgravity by deploying a protein-rich solution from a syringe, forming a drop in which crystallization can occur with the proper degree of supersaturation. Drop stability is critical to the success of this research, due to the large drop sizes which can be achieved in space. In order to determine the type of syringe tips most suitable to support these large drops, tests were performed during brief periods of weightlessness onboard the NASA KC-135 low-gravity simulation aircraft. The drops were analyzed using three simple models in which the samples were approximated by modified pendulum and spring systems. It was concluded that the higher frequency systems were the most stable, indicating that of the syringes utilized, a disk-shaped configuration provided the most stable environment of low-gravity protein crystal growth.

Owen, Robert B.↗

NASA NTRS1987

Control of electroosmosis in coated quartz capillaries

The effectiveness of various coatings for controlling the electroosmotic fluid flow that hinders electrophoretic processes is studied using analytical particle microelectrophoresis. The mobilities of 2-micron diameter glass and polystyrene latex spheres (exhibiting both negative and zero effective surface charge) were measured in 2-mm diameter quartz capillaries filled with NaCl solutions within the 3.5-7.8 pH range. It is found that capillary inner surface coatings using 5000 molecular weight (or higher) poly(ethylene glycol): significantly reduced electroosmosis within the selected pH range, were stable for long time periods, and appeared to be more effective than dextran, methylcellulose, or silane coatings.

Herren, Blair J.↗

NASA NTRS1987

Continuous flow electrophoretic separation of proteins and cells from mammalian tissues

This paper describes an apparatus for continuous flow electrophoresis (CFE), designed to separate macromolecules and cells at conditions of microgravity. In this CFE, buffer flows upward in a 120-cm long flow chamber, which is 16-cm wide x 3.0-mm thick in the microgravity version (and 6-cm wide x 1.5-mm thick in the unit-gravity laboratory version). Ovalbumin and rat serum albumin were separated in space (flight STS-4) with the same resolution of the two proteins achieved at 25 percent total w/v concentration that was obtained in the laboratory at 0.2 percent w/v concentration. Rat anterior pituitary cells, cultured human embryonic kidney cells, and canine Langerhans cells were separated into subpopulations (flight STS-8) more effectively than in unit gravity, with comparable resolution having been achieved at 100 times the concentration possible on earth.

Hymer, W. C.↗