Expression, purification and osteogenic bioactivity of recombinant human BMP-4, -9, -10, -11 and -14

Abstract

Bone morphogenetic proteins (BMPs) are cytokines from the TGF-beta superfamily, with important roles during embryonic development and in the induction of bone and cartilage tissue differentiation in the adult body. In this contribution, we report the expression of recombinant human BMP-4, BMP-9, BMP-10, BMP-11 (or growth differentiation factor-11, GDF-11) and BMP-14 (GDF-5), using Escherichia coli pET-25b vector. BMPs were overexpressed, purified by affinity his-tag chromatography and shown to induce the expression of early markers of bone differentiation (e.g. smad-1, smad-5, runx2/cbfa1, dlx5, osterix, osteopontin, bone sialoprotein and alkaline phosphatase) in C2C12 cells and in human adipose stem cells. The described approach is a promising method for producing large amounts of different recombinant BMPs that show potential for novel biomedical applications.

The Protein Kinase 2 Inhibitor CX-4945 Regulates Osteoclast and Osteoblast Differentiation In Vitro

“…The enhancing effect of CX-4945 on the induction of ALP was confirmed by qRT-PCR analysis (Fig. 3C), and moreover, the BMP-2-induced mRNA expression of osteogenic BMP-4 was also significantly enhanced by CX-4945 at 10 μM (Bessa et al, 2009). …”

Silk fibroin microparticles as carriers for delivery of human recombinant BMPs. Physical characterization and drug release

“…In fact, as we have observed the silk fibroin microparticles exhibited a slow rate of degradation. The release amounts during this phase were such that they could meet the concentrations of growth factor required in most in vitro and in vivo applications (in the range of few hundreds of nanograms to a few micrograms), by choosing a specific amount of BMP during the loading process (Kenley et al, 1994;Hosseinkhani et al, 2007;Wei et al, 2007;Bessa et al, 2009).…”

“…Human BMP-2, expressed in Chinese Hamster Ovary (CHO) cells, was purchased from Wyeth, UK. Human BMP-9 and BMP-14 were cloned and expressed in Escherichia coli, purified by histidine-tag affinity chromatography and freeze-dried, as described elsewhere (Bessa et al, 2009). All chemicals were of analytical grade and used as received.…”

How to use BMP-2 for clinical applications? A review on pros and cons of existing delivery strategies

“…Nevertheless, addition of fusion tags was in context of BMPs employed mostly for protein purification (histag [91], maltose binding protein [92]), or detection and enrichment purposes [93,94], and only in a few cases designed to enable a more permanent immobilization on scaffolds. Insertion of any kind of tag within the mature part of thr BMP sequence is strongly restricted to the N-terminus, which is caused by the buried architecture of its C-terminal end.…”