Chemistry: molecular biology and microbiology – Maintaining blood or sperm in a physiologically active state...
Patent
1992-09-11
1998-06-02
Ulm, John
Chemistry: molecular biology and microbiology
Maintaining blood or sperm in a physiologically active state...
424129, 435260, A01N 102, C12N 104
Patent
active
057597740
DESCRIPTION:
BRIEF SUMMARY
The present invention is directed to a method for detecting in vitro the presence or absence of circulating antibody types in a plasma, serum, antiserum or hypodermal fluid using diagnostic reagents comprising dried or lyophilized cells or cell-like materials.
BACKGROUND OF THE INVENTION
For the use of blood in transfusions it is important to test for cell-antibody incompatibility to ascertain properly, for example the ABO/Rh compatibility, since improperly matched transfusions can lead to death from mismatched blood. In addition to the conventional ABO/Rh typing, there is also a series of minor red cell surface antigens (approximately 400 or more are known) which can cause immune sensitization and alloimmune reactions if the patient is repeatedly exposed to these antigens. It is therefore important to have readily available blood typing kits which can be used to test donor blood and to test the serum of potential recipients. There are generally two varieties of blood typing kits: (1) a direct typing kit containing packaged known antisera to test agglutination behavior of sample red cells from donated blood cell units; and (2) "reverse typing" kits which contain known red cell standards used to test samples of plasma from potential recipients. The present invention is particularly applicable for use in the "reverse typing" kits for diagnosing blood samples.
Available "reverse typing" kits for clinical diagnostic use for typing blood comprise vials (usually from 2 to 30 ml each) of fresh human red blood cells of a known type suspended in a preservative medium called Alsever's solution, a solution of dextrose/sodium citrate, sodium chloride and an antibiotic, such as aureomycin, in water. However, suspensions of cells in Alsever's solution are generally limited to about a seven week refrigerated shelf life. The cells cannot be stored frozen in an Alsever's suspension. This relatively short shelf life poses a problem particularly for the hospital blood bank and the users since a typical manufacturing operation of an Alsever's suspension can take as long as three to four weeks from the time of donation to the time of regulatory release of the inventory for sale. This leaves the hospital blood bank and user only an effective three to four week shelf life before the kit must be destroyed.
Furthermore hospitals must also keep samples of rare type cell standards on hand (such as AB- and rare genotype standards) which are more expensive and difficult to acquire than commonly available cell standards. These must be replaced within the three to four week effective shelf life if they are not used.
Even during the seven weeks of refrigeration of red blood cells in Alsever's suspensions, key metabolites such as ATP, are depleted and slow cell lysis occurs. There is therefore a need for providing diagnostic kits for blood typing which have longer refrigerated (or room temperature) storage lives than the refrigerated Alsever's red cell suspensions.
The present invention is useful for, but not limited to, methods for blood grouping utilizing solid surfaces. For example, a solid surface capable of supporting an immunological reaction is provided on which cells or cell-like materials containing known antigens are attached or absorbed onto the surface. This surface containing the cells or cell-like materials may then be lyophilized or dried and stored until use. After rehydration, the surface may be contacted with an unknown blood component which may contain unknown antibodies specific for the known antigen previously attached, in which case, an immune reaction will occur. A monolayer of red blood cells containing the known antigen which was first attached to the solid surface may then be optionally activated by treating with a proteolytic enzyme. Alternatively, cells can be pretreated before attachment. The protease bromelain is known to enhance certain red blood cell antigen reactions with antibodies against these antigens. This known activated cell layer may then be brought into contact with the unknown blood component,
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Cho Miller
Galle Richard F.
Goodrich, Jr. Raymond P.
Hackett Roger W.
Olson Jon A.
Cobe Laboratories, Inc.
Ulm John
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