A flow cytometry-based method for detecting the cleavage of the extracellular segment of the platelet receptor gpiba

A technology of flow cytometry and platelets, applied in the field of medical biology, can solve the problems of high background value of ELISA, high level of soluble fragments, lower reliability and stability of test results, etc., and achieve high reliability and good stability
CN107340225BActive Publication Date: 2019-07-05XUZHOU MEDICAL UNIV
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
XUZHOU MEDICAL UNIV
Publication Date
2019-07-05

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Abstract

The invention discloses a method for detecting the enzyme cleavage of the extracellular segment of platelet receptor GPIba based on flow cytometry. The specific steps are: (1) sample preparation and processing; (2) flow cytometry detection; (3) Weasel Software data analysis. The present invention uses two different fluorescently labeled antibodies, respectively anti-GPIba extracellular segment antibody and anti-GPIba intracellular segment antibody, and analyzes the relative value of the combination of the extracellular segment antibody and the intracellular segment antibody by flow cytometry, namely It can reflect the degree of cleavage of the extracellular segment of GPIba. The detection method of the invention does not depend on the change of the number of platelets, is suitable for the situation of reduced number of platelets, and has high reliability and good stability.
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Description

technical field

[0001] The invention belongs to the technical field of medical biology, and in particular relates to a flow cytometry-based method for detecting enzyme cleavage of the extracellular segment of platelet receptor GPIba. Background technique

[0002] The GPIb-IX-V complex is composed of GPIba, GPIbb, GPIX and GPV ( figure 1 ). GPIba is combined with GPIbb through disulfide bonds, and then connected with GPIX and GPV through non-covalent bonds. GPIba is the major ligand-binding subunit in the GPIb-IX-V complex, consisting of 626 amino acid residues. GPIba is a membrane-expressed molecule consisting of an extracellular segment, a transmembrane region and an intracellular segment. As the most important adhesion molecule of platelets, GPIba exerts a variety of biological functions by binding to corresponding ligands.

[0003] After platelet activation or some kind of stimulation, the extracellular segment of GPIba will undergo enzyme cleavage, which is mainly me...

Claims

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