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[使用抗膜糖蛋白IIb-IIIa单克隆抗体对血小板进行放射性同位素标记以及测量血小板在底物表面的黏附/聚集]

[The radioisotope labelling of thrombocytes using a monoclonal antibody against membrane glycoproteins IIb-IIIa and the measurement of thrombocyte adhesion/aggregation on the substrate surface].

作者信息

Rainkina T V, Khorets B A, Katsenovich E R, Mazurov A V

出版信息

Biull Eksp Biol Med. 1992 Nov;114(11):529-32.

PMID:1290834
Abstract

A new method for platelet labeling based on binding of monoclonal antibody to human platelets has been suggested in this study. Monoclonal antibody VM16a against membrane glycoproteins IIb-IIIa was labeled by 125I and then incubated with platelets. About 70% of added antibody was bound when it was used at the concentrations corresponding to the linear part of the concentration curve (0.5 and 1.0 micrograms/ml). Due to high efficiency of binding 125I-VM16a-labeled platelets were used for the measurement of adhesion/aggregation to the substrate in platelet-rich plasma without washing of the free label. Experiments with washed platelets double labeled with 51Cr and 125I-VM 6a showed high correlation between the data obtained with both labels. The method of platelet labeling has been applied for the assessment of drug action on platelet adhesion/aggregation. Measurements were performed in platelet-rich plasma and adhesion/aggregation was stimulated by ADP and analogue of thromboxane A2, U46619. It was shown/that antianginal drug trapidil strongly inhibited and antiatherogenic drug probucol did not affect platelet adhesion/aggregation stimulated by both agonists.

摘要

本研究提出了一种基于单克隆抗体与人血小板结合的血小板标记新方法。抗膜糖蛋白IIb-IIIa的单克隆抗体VM16a用125I标记,然后与血小板孵育。当以对应于浓度曲线线性部分的浓度(0.5和1.0微克/毫升)使用时,约70%添加的抗体被结合。由于125I-VM16a标记血小板的结合效率高,因此无需洗涤游离标记物,即可用于测量富含血小板血浆中血小板与底物的粘附/聚集。用51Cr和125I-VM 6a双重标记的洗涤血小板进行的实验表明,两种标记物获得的数据具有高度相关性。血小板标记方法已应用于评估药物对血小板粘附/聚集的作用。在富含血小板血浆中进行测量,并用ADP和血栓素A2类似物U46619刺激粘附/聚集。结果表明,抗心绞痛药物曲匹地尔强烈抑制,而抗动脉粥样硬化药物普罗布考不影响两种激动剂刺激的血小板粘附/聚集。

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