Large-scale production of recombinant mammalian factor H (FH)

A bio-manufacturing method for high-yield, cost-effective production
Biotechnology
Medical technologies
Therapeutics

A bio-manufacturing method for high-yield, cost-effective production of a recombinant version of mammalian FH for use as a therapeutic agent.

Application

  • Treatment of age-related macular degeneration (AMD)
  • Treatment or prevention of diseases in which inadequate complement regulation contributes to aetiology or symptoms

Development Status

  • Undergoing further development for a range of diagnostic and therapeutic uses

IP Status

  • Patent granted US8889374B2, US10378005B2 and EP2516464B1

Commercial Offerings

  • Technology available for licensing

Opportunity

Human factor H (FH), a 155-kilodalton serum glycoprotein that regulates the complement system, is a potential bio-therapeutic for a range of human chronic conditions, including dry age-related macular degeneration (AMD) in genetically defined subpopulations and several rare kidney diseases. Attempts to produce FH recombinantly in many diverse host cell types failed to produce the quantities of material that would be required for clinical applications.

The patented technology offers a platform for the manufacture of recombinant full-length mammalian FH as well as domain-deleted, or otherwise engineered, versions of the glycoprotein. Production can be undertaken in industrial-scale fermenters, generating high-quality and functionally competent material in a rapid, cost-effective and efficient manner, in quantities suitable for many applications, including therapeutic use.

Technology

The patented technology involves the intracellular expression of a codon-optimized nucleic acid sequence encoding a mammalian FH (including human FH) or an engineered version of mammalian FH (including ‘mini’ versions that have enhanced activity). Rather than conventional gene cloning and expression, the method is based on gene design and synthesis techniques.

The method for FH production has been optimised for Pichia pastoris cells, an organism that grows easily on inexpensive and simple media in fermenter vessels to exceptionally high cell densities and secretes FH into the growth media whereupon it is easy to capture and purify. Moreover, many strains of P. pastoris are available including strains that specialise in making large quantities of proteins that contain numerous disulfides (FH has 40 disulfide bonds) and strains that decorate glycoproteins with humanised glycans (FH has eight N-glycans). In addition to making FH from fungal (yeast) cells the patented technology also covers FH production in bacterial, insect or plant cells.

Benefits

  • A high yield and cost-effective process for the production of mammalian FH.
  • Production of therapeutically useful and economically viable quantities of FH.
  • The potential to produce, in large quantities, versions of the glycoprotein with selected domains deleted or with low immunogenicity and extended in vivo half-lives.
  • Recombinantly generated FH and its engineered versions could be of high therapeutic value in countering complement activation and reducing complement-mediated damage to host tissues.

Publications

US8889374B2, US10378005B2, EP2516464B1 - Recombinant codon optimised factor H

Kerr H, Herbert AP, Makou E, Abramczyk D, Malik TH, Lomax-Browne H, Yang Y, Pappworth IY, Denton H, Richards A, Marchbank KJ, Pickering MC, Barlow PN. “Murine Factor H Co-Produced in Yeast With Protein Disulfide Isomerase Ameliorated C3 Dysregulation in Factor H-Deficient Mice”. Front Immunol. 2021. doi: 10.3389/fimmu.2021.681098.

Kerr H, Wong E, Makou E, Yang Y, Marchbank K, Kavanagh D, Richards A, Herbert AP, Barlow PN. “Disease-linked mutations in factor H reveal pivotal role of cofactor activity in self-surface-selective regulation of complement activation”. J Biol Chem. 2017. doi: 10.1074/jbc.M117.795088.

Christoph Q. Schmidt, Fern C. Slingsby, Anna Richards, and Paul N. Barlow. “Production of biologically active complement factor H in therapeutically useful quantities”, Protein Expression and Purification, 2011. doi: 10.1016/j.pep.2010.12.002.


Please note, the header image is purely illustrative. Source: Luchschen via GettyImages

Quote: TEC1102274

License this technology

Dr Maria Lopalco

Technology Transfer Manager
School of Chemistry
The Institute of Genetics and Cancer