Production of biologically active complement factor H in therapeutically useful quantities

Area of Science:

• Biochemistry
• Molecular Biology
• Biotechnology

Background:

• Human complement factor H (FH) is a critical regulator of the alternative complement pathway.
• FH gene mutations are linked to diseases like age-related macular degeneration and atypical hemolytic uremic syndrome.
• Current therapeutic strategies are limited by the reliance on plasma-derived FH.

Purpose of the Study:

• To develop a scalable method for producing recombinant human complement factor H (rFH).
• To ensure the functionality and therapeutic potential of rFH for treating complement-mediated diseases.

Main Methods:

• Recombinant FH (V62,Y402 variant) was expressed in Pichia pastoris, utilizing codon optimization and secretion via the α-factor peptide.
• Purification involved heparin-affinity and anion-exchange chromatography after endoglycosidase H digestion.
• Characterization included mass spectrometry, Western blot, and dynamic light scattering.

Main Results:

• Multiple 10s-of-milligrams of functional rFH were produced, significantly exceeding previous expression levels.
• rFH was confirmed as a single, non-aggregated species and demonstrated full biochemical and biological activity.
• A glycan-devoid version of rFH was also successfully produced.

Conclusions:

• Pichia pastoris is a viable and highly efficient system for producing therapeutically relevant quantities of rFH.
• This recombinant production method offers economic viability and potential for engineering enhanced FH variants.