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Methods for creating agglomerates from colloidal particles
| Details |
Inventors: Ekenberg, Steven J.;
Assignee: Promega Corporation (Madison, WI)
Primary Examiner: Kunz; Gary L.
Assistant Examiner:
Attorney, Agent or Firm: Frenchick; Grady J. Stroud, Stroud, Willink, Thompson & Howard
Methods for controllably creating agglomerates from particles of colloidal dimensions. Creation of agglomerates permits physical isolation or separation of, e.g., unwanted cell debris from cleared cell lysate comprising target particles, by the deposition or creation of a blocking or masking layer. Chaotropic agents are utilized to enhance colloidal particle agglomeration. The resulting agglomerant or "floc" can be deposited, as needed, in a separation/isolation process to create a blocking or masking layer on an "as needed" basis. |
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DETAILED DESCRIPTION As is described above, this invention is preferably utilized with magnetic separation devices for separation of magnetic particles from nonmagnetic media utilizing a multi-well or multi-titer separation device.
While a 96-well separation device is disclosed, a separator with five or more wells can be used more effectively in a practice of this invention.
The present invention is particularly well-suited for use in separating biological substances of interest in various laboratory and clinical procedures involving biospecific affinity reactions.
Accordingly, the present invention will now be described in detail with respect to such endeavors.
However, those skilled in the art will appreciate that such a description of the invention is meant to be exemplary only and should not be viewed as limitative of the full scope thereof.
A preferred method in accordance with the present invention utilizes particles which are magnetically responsive and which comprise a receptor capable of binding the substance of interest in the test sample.
After the receptor binds the target substance, a magnetic separator can be used to remove the magnetic particles bound to the substance of interest from the test medium.
Biospecific affinity reactions may be employed in testing biological samples for the determination of a wide range of target substances.
Representative target substances include cells, cell components, cell subpopulations (both eucaryotic and procaryotic), bacteria, parasites, antigens, proteins, specific antibodies, specific biological factors, such as vitamins, viruses and specific nucleic acid sequences, e.
g.
, mRNA.
Thus, the magnetic separation aspect of this invention has application in cell separations for the analysis or isolation of cells including, by way of example: T-cells from a T-cell lymphoma cell line; B-cells from a B-cell lymphoma cell line; CD4 positive cells from leukocytes; and lymphocytes from leukocytes.
The methods in accordance with the invention may also be used for immunospecific isolation of monocytes, granulocytes and other cell types; removal of rare cells; depletion of natural killer cells; determination of reticulocytes; and assays for neutrophil function, e
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Liquid Purification 
