Direct Determination of Coagulation Factor IIa and Plasmin Activities for Monitoring of Thrombotic State

Author:

Zhang Junhua12,Zou Lihui1,Liu Chengyang1,Li Chuanbao3,Wang Meng3,Yang He4,Wang Yan4,Tan Zheng4,Li Hexin2,Su Fei2,Zou Tong5,Li Kang5,Wang Xiaogang6,Li Ying12,Han Bingqing12,Zhang Zhu7,Zhai Zhenguo7,Liu Xiangyi8,Xu Xiaomao4,Xiao Fei12

Affiliation:

1. The Key Laboratory of Geriatrics, Beijing Hospital, National Center of Gerontology, Chinese Academy of Medical Sciences, Beijing, PR China

2. Clinical Biobank, Beijing Hospital, National Center of Gerontology, Chinese Academy of Medical Sciences, Beijing, PR China

3. Department of Laboratory Medicine, Beijing Hospital, National Center of Gerontology, Chinese Academy of Medical Sciences, Beijing, PR China

4. Department of Respiratory and Critical Care Medicine, Beijing Hospital, National Center of Gerontology, Chinese Academy of Medical Sciences, Beijing, PR China

5. Department of Cardiology, Beijing Hospital, National Center of Gerontology, Chinese Academy of Medical Sciences, Beijing, PR China

6. Department of Traditional Chinese Medicine, Beijing Hospital, National Center of Gerontology, Chinese Academy of Medical Sciences, Beijing, PR China

7. Department of Respiratory and Critical Care Medicine, The China-Japan Friendship Hospital, Beijing, PR China

8. Department of Clinical Laboratory, Beijing Tongren Hospital, Capital Medical University, Beijing, PR China

Abstract

Abstract Background Current laboratory examinations for hypercoagulable diseases focus on the biomarker content of the activated coagulation cascade and fibrinolytic system. Direct detection of physiologically important protease activities in blood remains a challenge. This study aims to develop a general approach that enables the determination of activities of crucial coagulation factors and plasmin in blood. Methods This assay is based on the proteolytic activation of an engineered zymogen of l-phenylalanine oxidase (proPAO), for which the specific blood protease cleavage sites were engineered between the inhibitory and activity domains of proPAO. Specific cleavage of the recombinant proenzyme leads to the activation of proPAO, followed by oxidation and oxygenation of l-phenylalanine, resulting in an increase of chromogenic production when coupled with the Trinder reaction. Results We applied this method to determine the activities of both coagulation factor IIa and plasmin in their physiologically relevant basal state and fully activated state in sodium citrate–anticoagulated plasma respectively. Factor IIa and plasmin activities could be dynamically monitored in patients with thrombotic disease who were taking oral anticoagulants and used for assessing the hypercoagulable state in pregnant women. Conclusions The high specificity, sensitivity, and stability of this novel assay not only makes it useful for determining clinically important protease activities in human blood and diagnosing thrombotic diseases but also provides a new way to monitor the effectiveness and safety of anticoagulant drugs.

Funder

National Key Research and Development Program of China

CAMS Innovation Fund for Medical Sciences

National Natural Science Foundation of China

Key Research Program for Health Care in China

Publisher

Oxford University Press (OUP)

Subject

General Medicine

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1. Research progress on biomarkers of pulmonary embolism;The Clinical Respiratory Journal;2021-08-03

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