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Binding ability of methylene blue with heparin dependent on its sulfate level rather than its sulfation location or basic saccharide structure

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Abstract

Methylene blue (MB) is one of the most common cationic dyes to detect heparin. As the sulfate residue presented in heparin was the main contributor to bind with MB, the UV performance of the MB with selectively desulfated heparin derivatives was investigated. It was found that the sulfate residue in different heparin analogues did not show the equal ability to attract MB binding. The stoichiometry of sulfate with MB among the heparin and derivatives was verified as a non-constant number. For the two selectively desulfated heparin derivatives: sulfate elimination at 6-O (6-OdeS) and N-acetylated heparin (N-deS-Acetyl), the MB to sulfate ratios were significantly higher than for heparin. For the not fully diminished sulfate at 2-O heparin derivative (2-OdeS), the MB-SO3- ratio of 2-OdeS was between 6-OdeS, N-deS-Acetlyl and heparin. Although in a distinct sulfation position, the MB-SO3- ratio of 6-OdeS and N-deS-Acetyl was almost equal, which agreed with the comparable total desulfation degree between 6-OdeS and N-deS-Acetyl. In addition, compared to heparin groups, the non-desulfated gs-HP showed no significantly different MB-SO3- ratio with heparin. The above results demonstrated that compared with the sulfate location and glycan composition of heparin, the content of sulfate was the most essential factor for the MB binding.

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Acknowledgments

We all gratefully acknowledge the financial support of the National Natural Science Foundation of China (Grant No. 82003647, 21807063), the Natural Science Foundation of Shandong Province (Grant No. ZR2019BH045), China Postdoctoral Science Foundation (Grant No. 2019M652307, 2020T130332) and Start up grant of Qingdao University of Science and Technology (Grant No. 210/010029032).

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Authors and Affiliations

  1. State Key Laboratory Base for Eco-Chemical Engineering in College of Chemical Engineering, Qingdao University of Science and Technology, Qingdao, 266042, China

    Shi-Xi Jia, Qiao-Na Chi, Yuanyuan Zhang, Tao Liu, Fanye Wang & Shan-Shan Du

  2. Analyses and testing center, Qingdao University of Science and Technology, Qingdao, 266042, China

    Xinhui Kou

  3. Department of Medical Chemistry, School of Pharmacy, Qingdao University, Qingdao, 266021, China

    Yun-Kun Qi

  4. Department of Genetics and Cell Biology, Basic Medical College, Qingdao University, Qingdao, 266071, China

    Shan-Shan Du

  5. MOE Key Laboratory of Industrial Biocatalysis, Institute of Biochemical Engineering, Department of Chemical Engineering, Tsinghua University, Beijing, 100084, China

    Xin-Hui Xing

  6. Centre for Synthetic and Systems Biology, Tsinghua University, Beijing, 100084, China

    Xin-Hui Xing

  7. Institute of Biopharmaceutical and Health Engineering, Tsinghua Shenzhen International Graduate School, Shenzhen, 518055, China

    Xin-Hui Xing

  8. Institute of Biomedical Health Technology and Engineering, Shenzhen Bay Laboratory, Shenzhen, China

    Xin-Hui Xing

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  1. Shi-Xi Jia
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Correspondence to Shan-Shan Du.

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Jia, SX., Chi, QN., Zhang, Y. et al. Binding ability of methylene blue with heparin dependent on its sulfate level rather than its sulfation location or basic saccharide structure. Glycoconj J 38, 551–560 (2021). https://doi.org/10.1007/s10719-021-10010-2

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