Development of a method to analyze the complexes of enoxaparin and platelet factor 4 with size-exclusion chromatography

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Authors
Wu, Fangxia
Dong, Kai
Zhu, Meng
Zhang, Qinghua
Xie, Bingying
Li, Duxin
Gan, Hao
Linhardt, Robert J.
Zhang, Zhenqing
Issue Date
2019-02-05
Type
Article
Language
ENG
Keywords
Biology , Chemistry and chemical biology , Chemical and biological engineering , Biomedical engineering
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Abstract
Heparin, a highly sulfated glycosaminoglycan, has been used as a clinical anticoagulant over 80 years. However, heparin-induced thrombocytopenia and thrombosis (HITT) is a serious side effect of heparin therapy, resulting in relatively high risk of amputation and even death. HITT is caused by forming of complexes between heparin and platelet factor 4 (PF4). Enoxaparin, one of the most commonly used low molecular weight heparin (LMWH), were developed in 1980's. The lower molecular weight of enoxaparin reduces the risk of HITT by binding to less PF4. To detect the binding capacity between enoxaparin and PF4 could be an effect way to control this risk before it goes to patients. In this work, a size exclusion chromatography (SEC) method was developed to analyze the patterns of complexes formed between PF4 and enoxaparin. The chromatographic condition was optimized to separate PF4, enoxaparin, ultra-large complexes and small complexes. The linearity and stability of this method were confirmed. The impacts of PF4/enoxaparin mixture ratios and incubation time on the forming complexes were investigated. Four enoxaparin samples were analyzed with this method to verify its practicability. It is a robust, accurate and practicable method, and provides an easy way to monitor the capacity of enoxaparin forming complexes with PF4, suggesting the HITT related quality of enoxaparin.
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Journal of Pharmaceutical and Biomedical Analalysis 164, 668-671
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Full Citation
Development of a method to analyze the complexes of enoxaparin and platelet factor 4 with size-exclusion chromatography, F. Wu, K. Dong, M. Zhu, Q. Zhang, B. Xie, D. Li, H. Gan, R.J. Linhardt, Z. Zhang, Journal of Pharmaceutical and Biomedical Analalysis 164, 668-671, 2019.
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1873264X
7317085
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