Overproduction of recombinant human bone morphogenetic protein-7 in Chinese hamster ovary cells

Area of Science:

• Biotechnology
• Protein Engineering
• Cell Line Development

Background:

• Bone Morphogenetic Protein-7 (BMP-7) is a crucial growth factor with diverse roles in cellular processes.
• BMP-7 has demonstrated therapeutic potential for various diseases, including bone metastasis and developmental disorders.
• Efficient and scalable production of functional BMP-7 is essential for its clinical applications.

Purpose of the Study:

• To establish a stable Chinese hamster ovary (CHO) cell line for high-level expression of recombinant human BMP-7 (rhBMP-7).
• To optimize the fed-batch culture process for maximizing rhBMP-7 production.
• To purify and characterize the functional activity of the produced rhBMP-7.

Main Methods:

• Development of a stable CHO cell line overexpressing rhBMP-7.
• Optimization of a 14-day fed-batch culture in a 7.5-L bioreactor using chemically defined medium.
• Two-step chromatographic purification of rhBMP-7 from culture supernatant.
• Functional assessment of purified rhBMP-7 using MC3T3-E1 cell proliferation assays.

Main Results:

• The engineered CHO cell line produced over 188 mg/L of rhBMP-7 in a 14-day fed-batch culture.
• rhBMP-7 was purified to >95% purity with approximately 55% recovery using a two-step chromatographic method.
• The purified rhBMP-7 exhibited significant biological activity, with a half-maximal effective concentration (EC50) of 28.31 ng/mL for MC3T3-E1 cell proliferation.

Conclusions:

• A robust and scalable method for producing high-titer, functional rhBMP-7 using a stable CHO cell line has been established.
• The developed production and purification process yields high-purity, biologically active rhBMP-7 suitable for further therapeutic development.
• This study provides a foundation for the large-scale manufacturing of rhBMP-7 for potential clinical applications.