Chemistry: electrical and wave energy – Apparatus – Coating – forming or etching by sputtering
Patent
1993-06-09
1995-02-07
Niebling, John
Chemistry: electrical and wave energy
Apparatus
Coating, forming or etching by sputtering
2041823, B01D 6146
Patent
active
053873254
DESCRIPTION:
BRIEF SUMMARY
BACKGROUND OF THE INVENTION
1. Field of the Invention
This invention relates generally to methods and a system for the separation and resolution of molecular components, including synthetic, organic or biologic particles, from a mixture of such components. More particularly, the invention pertains to a multilayer molecular filtration system comprised of discontinuous and nonsequential, gelous, semifluid or fluid, chromatographic or electrophoretic mediae placed on or inside or around different and presently known and used mechanical vectors, facilitating a movement of polar particles with various molecular characteristics and related methods thereof.
2. Description of Related Art
The use of sieving gels for the separation of organical and/or biological particles is well-known and used in the electrophoresis art from mid-fifties of this century. However, the presently known methods, based on sieving effects demonstrated throughout the years, have serious disadvantages. The most apparent is the inability to separate, satisfactorily, the high and very high molecular particles, e.g., molecular weights greater than 650,000 daltons.
The sieving gels which are most often used for analytical and scientific purposes include the following: cellulose acetate, starch gels, agar-gels, agarose-gels, gels of agarose derivatives, polyacrylamide gels, gradient polyacrylamide gels of different gradients (from 2% to 27% of PAA and 4% to 30% of PAA), pore gradient polyacrylamide gels, mixture of, or combinations of polyacrylamide gels and different derivatives of agarose, as well as any of the above gels with addition of protein-complexes-forming variants (SDS), urea, or emulators (TRITONS), or glass adhesives, etc.
All the above-mentioned gels and combinations thereof are known to be used only in continuous forms (e.g., nongradient mediae), or continuous gradients (e.g., gradient mediae). For example, the most often employed gelous mediae are the agarose-gels. Although gels of different concentrations of purified agarose are utilized for specific purposes (e.g., 1%, 11/2%, 2%, 21/2%, etc.), all are known as "continuous" gels. In comparison, the "gradient" gels comprise gels (usually polyacrylamidic) of increasing concentrations of acrylamide monomer, as shown by the formula where T equals the percent of monomers concentration: ##EQU1## These gels are known to be used in different ranges of their gradients (for instance from 3-7%, or 2-5%, or 2-16%, or 2-27%), but these gradients are known to be always sequentially assorted (e.g., in 3-4-5-6-7%, or 2-3-4-5%, or in 2-3-4-5-6-7-8-9-10-11-12-13-14-15-16% fashion, etc.).
It is noted that the literature discloses only few devices of similar arrangement of two electrophoretic gels from which two gels designated (A,B) were called "discontinuous", and one designated (C) was described as a gel with a "stepped gradient". However, none of the above arrangements suggest a variable multilayer system of distinct electrophoretic gels, forming specific molecular sieves characterized by retention gradients for elected groups of polar biomolecules.
The above designated devices (A, B and C) only comprise two (2) layers of gels wherein the first layer serves primarily as a mechanical, and not as an analytical means (e.g., delivery of the biological sample to an analytical gel (A), or to clean the analyzed biological sample from interfering components (B), or to unspecified improvement of desired analytical procedures (C)). In addition, certain ones, called "technical" gels are composed of components which are substantially smaller, than are the main, analytical gels (A, B, C).
U.S. Pat. No. 4,306,956 to De Castro describes the first of the two-layer systems designated (A). This patent is directed to the use of thymol blue, phenol red, o-cresol red, orange G, m-cresol purple and mixtures, as a tracking dye in an SDS-PAGE electrophoresis process. The patent describes system designated (A) as follows--"Often in discontinuous sodium-dodecyl dodecyl sulfate polyacrylamide electrophoretic gels (U.S.
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Allen, Ph.D., Pharmazeutisches Institut der FU,D-1 Berlin 33, Konigin-Luise-Strasse 2-4, Germany, "Electrophoretic Separation of Pre-Stained Lipoproteins on Polyacrylamide Gel Slabs and their Relationship to Other Plasma Proteins". Oct. 6-7, 1972.
Niebling John
Wong Edna
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