E. coli platform for expression of low-cost malaria fusion proteins
Award last edited on: 1/31/2024

Sponsored Program
Awarding Agency
Total Award Amount
Award Phase
Solicitation Topic Code
Principal Investigator
Andrew Lees

Company Information

Fina Biosolutions LLC

9430 Key West Avenue Suite 200
Rockville, MD 20850
   (877) 346-2246
Location: Single
Congr. District: 08
County: Montgomery

Phase I

Contract Number: 2024
Start Date: ----    Completed: 1/1/2024
Phase I year
Phase I Amount
The development of a malaria vaccine is "one of the most important researchprojects in public health" (CDC website). In 2021, there were 250 million malaria cases andmore than 600,000 deaths, with most deaths in children under 5. About 95% of malariadeaths are in the sub-Sahara, home to some of the world's poorest countries. Even with thefirst malaria vaccine approved and more vaccines in the pipeline, there is a need for malariavaccines that are both clinically effective and affordable. Malaria vaccine antigens arechallenging to express in the needed quantities and at the needed price. Furthermore, asmalaria antigens are generally poorly immunogenic, they are often chemically conjugated toa carrier protein to make nanoparticle vaccines. This SBIR proposal is directed to makingthese antigens easier and less expensive to manufacture. Our collaborators at Oxford University and the NIH Laboratory of MalariaImmunology and Vaccinology have identified domains of blood-stage and transmission-blocking malaria proteins which elicit blocking antibodies, but they have only been made inexpensive eukaryotic systems but have not been successfully made in low-cost bacteria likeE. coli. We have developed the Gor∆ E. coli strain for producing difficult-to-express proteins.Gor∆ has an oxidative cytoplasm and can make soluble, correctly folded disulfide bondproteins in the cytoplasm at high yields. We have used this strain to make multi-gram/L ofsoluble CRM197, a widely used vaccine carrier protein. By creating genetic fusions withCRM197 as the partner, we could express proteins that otherwise could not be made in E.coli. In this SBIR, we will: (1) Use Gor∆ to make CRM197 fusion proteins with (a) a domain ofblood-stage antigen, RH5, and (b) domains of transmission-blocking antigens, Pfs230 andPfs48/45; (2) Confirm proper folding using conformationally-dependent monoclonalantibodies as well as perform biophysical analysis for correct MW, sequence, and disulfidebonding, and (3) Synthesize chemical conjugates of the fusion proteins and compare theirimmunogenicity to the unconjugated proteins. Antisera will be evaluated for blood-stageinhibition using the growth inhibition assay and transmission-blocking activity with thestandard membrane feeding assay. This SBIR will allow us to demonstrate the utility of ourGor∆ E. coli strain to manufacture affordable malaria vaccine antigens as well as an array ofother vaccine proteins.

Public Health Relevance Statement:
Malaria is caused by a parasite with a complex lifecycle, including a blood stage in which the parasite binds and enters red blood cells: A mosquito acquires malaria from the blood during feeding and transmits the parasite on biting its next victim. Transmission-blocking vaccines induce antibodies that will block parasite passage through the mosquito; both transmission-blocking and blood-stage vaccine antigens have been identified, but, to date, they have only been produced in expensive eukaryotic systems. We propose to produce these antigens as genetic fusions in our engineered E. coli strain, which can make soluble, complex proteins in the cytoplasm at high levels, allowing us to manufacture a low-cost malaria vaccine targeting multiple elements of the malaria parasite lifecycle.

Project Terms:
<0-11 years old>

Phase II

Contract Number: 1R43AI181078-01
Start Date: 12/31/2024    Completed: 00/00/00
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