Method of separating particles from a filter

Details Method of separating particles from a filter Method of separating particles from a filter

1995-10-03

1998-11-03

Reifsnyder, David A.

Liquid purification or separation

Processes

Liquid/liquid solvent or colloidal extraction or diffusing...

210662, 210772, 210782, 210785, 210787, 21032169, 21050021, 356 39, 422101, 436178, B01D 6100

Patent

active

058303596

DESCRIPTION:

BRIEF SUMMARY
BACKGROUND OF THE INVENTION

1. Field of the Invention
The invention relates to a method of separating particles from a filter material on which they are retained by adsorption and a method of recovering particles from fluids containing them. More particularly, the invention relates to a method for rapidly and selectively recovering biological particles, such as leucocytes and platelets, in a form in which they can be used for many different purposes.
2. Discussion of Prior Art
There are many fluids in which leucocytes may occur: whole blood, blood components, milk, colostrum, urine, tissue and tumour disaggregates and exudates, lymph, ascites fluid, cerebrospinal fluid, bile, peritoneal fluid, synovial fluid, seminal fluid, lacrimal fluid, interstitial fluid, hemolymph, saliva, tears, mantle fluid, bone marrow, coelomic fluid, glandular secretions, bronchoalveolar lavage, alveolar fluid, fluid from organs and tissues in culture, pathological discharges, discharges, mucus, pus and odematous fluid that accumulates in any space within a living organism.
Much information can be obtained from measuring, for example, the production by leucocytes of free radicals and other reactive oxygen containing species and of the enzymes produced and sometimes released by leucocytes when they become activated. These can be measured by adding a suitable luminogenic material, e.g., Pholasin (Registered Trade Mark), which will emit light in the presence of free radicals, certain oxidants, some reactive oxygen species and certain enzymes and any light emitted would be a measure of the activity of the leucocytes. Many other tests, including non-luminogenic tests are also performed on leucocytes. However, it is often essential if such tests are to be performed accurately, that the large majority of other cells, such as red blood cells, which may be present in the fluid from which the leucocytes were harvested are removed. It is also important in many cases to remove the plasma (or serum) and other fluid in which the leucocytes occur before carrying out many tests as components in the fluid may interfere with the performance of many tests.
One approach to the problem of potential interference from plasma components, such as complement, is to dilute a sample of whole blood at least 1:500. However, this procedure reduces at the same time the number of cells present in the sample by the dilution factor and, therefore, any measurable signal is also correspondingly reduced. As leucocytes, in whole blood, are in relatively small numbers compared to the red blood cells, any tests requiring large number of leucocytes isolated from red blood cells, or tests requiring the extraction of specific components from leucocytes, would still not be possible using samples of diluted whole blood.
There is a recurrent need selectively to separate biological particles, such as leucocytes or platelets, from blood, for example, for subsequent analysis. There is also a need selectively to separate leucocytes from other body fluids, for example seminal fluid, in order to obtain a pure sample of sperm which are to be used in subsequent fertility tests.
There are known methods for separating biological particles, such as leucocytes, from blood, for example, for subsequent analysis or even for treatment to insert a new gene, tag a radioactive label to the cell or other such procedure which involves removing leucocytes and then returning such modified cells to the whole organism or to an isolated organ or tissue. Such methods include multistep procedures involving sedimentation with dextran, followed by separation on density gradients. These procedures involve centrifugation, mixing, incubating and sometimes lysis of unwanted red blood cells. They take a number of hours to complete, involve skilled operatives and subject the cells to uncontrollable variables which may inadvertently affect their subsequent response to analytical procedures (Boynum, A. (1968) "Isolation of mononuclear cells and granulocytes from human blood". Scand. J. Clin. Lab. Invest. 21, Sup

REFERENCES:
patent: 4330410 (1982-05-01), Takenaka et al.
patent: 5240613 (1993-08-01), Tsuchitani et al.
patent: 5252228 (1993-10-01), Stokes et al.

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