Department of Biochemistry and Molecular Biology, Michigan State University, 603 Wilson Road, East Lansing, 48824, MI, USA.
Regeneron Pharma, Tarrytown, NY, 10591, USA.
Mol Biotechnol. 2022 Feb;64(2):156-170. doi: 10.1007/s12033-021-00403-x. Epub 2021 Sep 22.
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.
重组人 BMP-4 生长因子 (GF) 在再生骨骼的治疗和细胞培养补充方面具有重要的商业潜力。然而，由于大规模表达和纯化人 BMP-4 GF 的尝试证明具有挑战性，其商业用途受到限制。我们已经建立了一种从中国仓鼠卵巢细胞培养物中提取 GF 部分的新方法，从细胞外基质或细胞沉淀部分获得大量纯净且具有生物活性的 BMP-4 GF。这种方法使产量比从 CM 中纯化的 BMP-4 GF 增加了约 100 倍。两种级分的分子活性无法区分。我们进一步分析了 BMP-4 GF 与蛋白聚糖肝素的结合，并表明 N 端碱性序列对于这种相互作用是必需的。综上所述，这些结果为 BMP-4 的纯化、定位和肝素结合提供了新的见解，这对其生物加工和生物学功能具有重要意义。
