Biologically active recombinant human complement factor H: synthesis and secretion by the baculovirus system

Area of Science:

• Biochemistry
• Molecular Biology
• Immunology

Background:

• Human complement factor H (hCFH) is a critical regulator of the complement system, involved in innate immunity and preventing autoimmune diseases.
• Deficiency or dysfunction of hCFH is linked to various pathologies, including age-related macular degeneration and atypical hemolytic uremic syndrome.
• Efficient production of biologically active hCFH is essential for research and therapeutic development.

Purpose of the Study:

• To establish a heterologous expression system for producing biologically active human complement factor H (hCFH).
• To clone and express the complete cDNA encoding hCFH in a scalable and efficient manner.
• To characterize the biological activity and yield of the recombinant hCFH protein.

Main Methods:

• Cloning of a 4.3 kb complete cDNA encoding human complement factor H (hCFH) into the pVL1393 baculovirus expression vector.
• Transfection of the recombinant vector into Spodoptera frugiperda Sf9 insect cells for protein expression.
• Purification and biological activity assays of the secreted recombinant hCFH protein.

Main Results:

• Successful cloning and expression of hCFH in a baculovirus-insect cell system.
• Secretion of a biologically active 140-kDa hCFH protein into the culture medium.
• Achieved a high yield of over 5 mg/liter with 92% +/- 15% biological activity.

Conclusions:

• This study reports the first successful synthesis of biologically active recombinant human complement factor H (hCFH) in a heterologous system.
• The developed baculovirus expression system provides a scalable and efficient method for producing functional hCFH.
• This recombinant hCFH is suitable for further functional studies and potential therapeutic applications.