Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving antigen-antibody binding – specific binding protein...
Patent
1994-02-07
1998-07-07
Stucker, Jeffrey
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving antigen-antibody binding, specific binding protein...
435 72, 435 724, 436 52, 436512, 436518, G01N 3353
Patent
active
057766975
DESCRIPTION:
BRIEF SUMMARY
TECHNICAL FIELD
The present invention relates to a process for the characterization or determination of the amount of mammalian cells according to the technique previously known per se where antibodies are utilized for said purpose. Up to now antibodies prepared in a mammal, especially rabbit, have been utilized for such purposes, but the present invention relates to the utilization of other types of antibodies in these connections, whereby disadvantages in connection with the prior art are eliminated, even in such a way that the invention enables such characterizations or determinations which would otherwise be troublesome or even impossible.
BACKGROUND OF THE INVENTION
Although the invention is generally applicable to the characterization or determination of many different types of mammalian cells, which will be more specifically exemplified below, it is of a very great value in connection with thrombocytes (blood platelets), especially with reference to the central parts of the thrombocyte functions in different connections. Therefore, a background of the invention will be given with reference to thrombocytes, as the problems to be solved by the invention and the advantages thereof will be most obvious thereby. However, corresponding or similar advantages can be achieved in the determination of other mammalian cells.
Thrombocytes have a central part of the haemostasis. It is well known that low thrombocyte numbers in the blood give rise to bleeding complications. There are also data which indicate that an increased number of activated thrombocytes and/or a reduced activation threshold may have an importance for the development of thrombi. Also such diseases as preeclampsia (a syndrome of pregnants with the symptoms of high blood pressure and urine protein) are associated with such alterations of the normal thrombocyte functions. Examinations of the thrombocyte functions can also be expected to become of great importance for decisions as to which patients should receive thrombocyte concentrates.
Haemostasis requires aggregation of thrombocytes. However, said aggregation can take place only if the thrombocytes will expose receptors for fibrinogen on their surfaces. In their condition of rest the thrombocytes have a minor capability of binding fibrinogen and then they can not either form aggregates which are the primary haemostatic plug (the first blocking of a vascular lesion) or part of a thrombosis (a clot). The activation of thrombocytes means that the thrombocytes will get a significantly increased capability of binding fibrinogen.
In other words there is a great demand of determining, in the blood of a patient, the amount or portion of the thrombocytes which are activated, i.e. which are highly capable of binding fibrinogen. However, those routine methods which have been utilized for testing the thrombocyte function, e.g. bleeding time and thrombocyte aggregation, are complex and insensitive and merely give a non-specific information about the function. For instance, they will give no picture of the discrete or separate thrombocytes but merely an average value. Furthermore, there are methods for determining proteins which are liberated from the thrombocytes, but the results thereof are unreliable as they are influenced inter alia by the metabolism of the patient.
In recent years fluorescence activated flow cytometry has, however, made it possible to develop new assays of thrombocyte functions. In this context laser beams are utilized which illuminate cells in a passing liquid jet. The light scattering gives information about the sizes of the cells and their structures. Further information will be obtained by the addition of antibodies or other substances which have been linked to different substances which when illuminated are caused to produce light (fluorescence). These methods are rapid and sensitive and can give a specific information about thrombocyte functions. Moreover, it is possible to study each thrombocyte separately and not only an average of all thrombocytes. For an assay a few micro
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Larsson Anders
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