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BMP-4 Extraction from Extracellular Matrix and Analysis of Heparin-Binding Properties

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Abstract

Recombinant human BMP-4 growth factor (GF) has significant commercial potential as therapeutic for regenerating bone and as cell culture supplement. However, its commercial utility has been limited as large-scale attempts to express and purify human BMP-4 GF have proved challenging. We have established a novel approach to obtain significant quantities of pure and bioactive BMP-4 GF from Chinese hamster ovary cell cultures by extracting the GF moiety from the extracellular matrix or cell pellet fraction. This approach increased yields approximately one 100-fold over BMP-4 GF purified from CM. The molecular activities of the two fractions are indistinguishable. We further analyzed binding of BMP-4 GF to the proteoglycan Heparin and showed that an N-terminal basic sequence is essential for this interaction. Taken together, these results provide novel insights into the purification, localization, and Heparin binding of human BMP-4 that have implications for its bioprocessing and biological function.

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Funding

This work was funded by NIH Grant R01 GM121499 to EMH.

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Author notes

  1. Senem Aykul

    Present address: Regeneron Pharma, Tarrytown, NY, 10591, USA

Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, Michigan State University, 603 Wilson Road, East Lansing, 48824, MI, USA

    Senem Aykul, Jordan Maust & Erik Martinez-Hackert

Authors
  1. Senem Aykul
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  2. Jordan Maust
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Contributions

SA and JM designed, performed, and analyzed experiments; EMH designed and analyzed experiments; EMH wrote manuscript.

Corresponding author

Correspondence to Erik Martinez-Hackert.

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Conflict of interest

EMH holds shares of Acceleron Pharma. EMH is a shareholder and co-founder of Advertent Biotherapeutics. SA is an employee and holds stock options of Regeneron Pharma. JM has no competing interests.

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Aykul, S., Maust, J. & Martinez-Hackert, E. BMP-4 Extraction from Extracellular Matrix and Analysis of Heparin-Binding Properties. Mol Biotechnol 64, 156–170 (2022). https://doi.org/10.1007/s12033-021-00403-x

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  • DOI: https://doi.org/10.1007/s12033-021-00403-x

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