Stock material or miscellaneous articles – Coated or structually defined flake – particle – cell – strand,... – Particulate matter
Reexamination Certificate
2001-05-02
2003-12-02
Barry, Chester T. (Department: 1724)
Stock material or miscellaneous articles
Coated or structually defined flake, particle, cell, strand,...
Particulate matter
C428S403000, C428S407000, C210S807000, C210S222000, C210S645000, C210S263000, C210S767000, C210S667000, C210S691000
Reexamination Certificate
active
06656587
ABSTRACT:
BACKGROUND OF THE INVENTION
1. Field of the Invention
The present invention relates to materials and methods for contacting a solution with a substrate and separating the substrate from the solution. More specifically, the present invention relates to composite micro-sized, magnetic particles for use in extracting desirable or undesirable components from a suspension or solution.
2. Description of the Related Art
Current methods for separating biological materials from impurities and/or suspending media employ the use of very fine magnetic particles that have coated thereon an active separating material. In this context, it is desirable that the magnetic materials possess at least two physical properties, namely, (1) a proper combined mass and size and (2) paramagnetic qualities. The former property will allow the particle to settle more slowly in suspension, thereby enhancing the particles' exposure to and interaction with the suspension. Thus, the smaller they are, the more slowly the particles will settle out according to Stokes Law and the effects of Brownian motion in the case of ultra small particles. The latter paramagnetism property allows the particles to be subsequently removed from the suspension by application of a magnetic field and for example, decanting off the suspending liquid. Since the particles are paramagnetic they will not have had induced residual magnetism and with the field removed, can be re-suspended in yet another recovery medium if necessary without clumping together.
Accordingly, current practice involves the use of very fine paramagnetic particles, consisting of iron oxide and silica composites, some of which are coated and others are mixtures. These particles typically are on the order of 10 to 100 nanometers, for example, and are suspended directly in a solution or suspension containing the nucleic acid or other molecules capable of being extracted from the suspension. Generally, these nano-sized particles contain a coating of an “active” material, that is, a material that has an affinity for a desired material already in suspension or solution. The coated magnetic particles are then separated from the suspension by application of a magnetic field.
Once the desired material in suspension has bound to the active material that is coated on the nano-sized particles, the particles are removed from the suspension. These bound materials can be removed by dissolution with reagents. However, these nano-sized particles often are too minute to separate completely from the suspension. Further, the high surface area of the fine particles increases their own susceptibility to dissolution as well, thus adding an impurity to the extracted media. Thus, a substantial concentration of these particles may remain in suspension and are lost in waste streams. Still further, undesirable clumping may occur when nucleic acid molecules attach to multiple magnetic particles, which are of comparable size, forming chains or large groups of the two. As a result, it is difficult to obtain desirable amounts of material that may have adhered to the particles. For the particles that actually are separated from suspension, multiple successive rinsing steps with extractive solutions are required.
Therefore, there is a present need for larger particles, for example, particles on the order of a sub-micron size to tens of microns, which would perform the function of material removal at high yield and be magnetically separable. By virtue of their size, micro-sized particles meeting these criteria could be separated from suspension more easily than nano-sized particles. Accordingly, the use of these particles would facilitate robotic manipulation of the separation process.
However, an increase in the diameter of these sphere-like particles disproportionately increases their mass, typically resulting in an increased rate of settling out of suspension. Further, agitation such as by stirring to maintain suspension may damage the delicate bio-substances. Hence, there is also a need for a gentle means to keep the particles suspended for times sufficient to allow the desired removal processes to take place.
SUMMARY OF THE INVENTION
Accordingly, it is an object of the invention to provide a micron-sized composite particle that is capable of interacting with a targeted material from solution, yet does not settle out of suspension at a rate typically associated with conventional micron-sized particles.
It is a further object of the invention to provide a micron-sized composite particle that is capable of isolating a targeted material from solution, yet does not settle out of suspension at a rate typically associated with conventional micron-sized particles.
It is, therefore, another object of the invention to provide a micron-sized composite substrate having (1) paramagnetic properties; (2) materials whose properties are designed to separate the desired substances from the suspension; and (3) to provide the buoyancy necessary to retard settling time for the extraction media to remove the desired substances.
These and other objects of the invention will become apparent upon reading the disclosure and teachings set forth herein.
In a compositional sense, the invention provides a composite material having an admixture of at least one buoyant particle, a variable blend of magnetic material that is susceptible to an induced magnetic field, and an active material. In one preferred embodiment, the above composite material is suitable for holding the composite in suspension in a fluid for a selected length of time and the active material is capable of adsorbing and/or reacting with at least one substance in the fluid and has a size on the order of about 10 &mgr;m to about 300 &mgr;m.
The individual components of the inventive composite material can be constructed in a number of ways. For instance, the variable blend of magnetic material can be chemically vapor deposited or wash-coated on the buoyant particle, and the active material can be chemically vapor deposited or applied via a sol gel process. In addition, the buoyant material may contain magnetic material incorporated therein, wherein the magnetic material is susceptible to an induced magnetic field.
A composite material of the invention can be used in conjunction with many different technologies. For instance, the composite material can be used to extract a biological material from a solution. The composite material also can be used to separate an impurity from a fluid.
In a methodological sense, the invention provides a method for extracting a biological material or impurity from a solution, including the steps of: providing a composite material separation medium containing one or more buoyant particles, a variable blend of magnetic material, and a material having an affinity for the biological material or said impurity; contacting the separation medium with a solution containing the biological material or impurity, wherein at least a portion of the biological material or impurity is bound to the material having an affinity therefor; removing the separation medium containing the bound biological material or impurity from the solution; and separating the bound biological material or impurity from the separation medium.
The present invention also includes a method of controlling the time of suspension of an active material in a fluid, containing the steps of: providing a composite material as described herein; contacting the composite material with a fluid in an amount sufficient to suspend the composite material, whereby the amount of time the active material is suspended depends on the overall density of the composite material in accordance with Stoke's Law.
REFERENCES:
patent: 2093454 (1937-09-01), Kistler
patent: 3117027 (1964-01-01), Lindlof et al.
patent: 3657119 (1972-04-01), Turbeville
patent: 3864124 (1975-02-01), Breton et al.
patent: 3897221 (1975-07-01), Salyer et al.
patent: 3970518 (1976-07-01), Giaever
patent: 4349456 (1982-09-01), Sowman
patent: 4454234 (1984-06-01), Czerlinski
patent: 4666854 (1987-05-01), Sugiura
Entezarian Majid
Johnson James R.
Barry Chester T.
Phillips Plastics Corporation
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