# Toxoid adjuvant CRM197 production in a stable reduced genome E. coli strain

> **NIH NIH R44** · SCARAB GENOMICS, LLC · 2021 · $1,000,000

## Abstract

Scarab Genomics has established a unique, tiny footprint, extended bacterial fermentation process enabled by
its reduced genome E. coli, for production of protein therapeutics, especially vaccine conjugates. The process
will massively reduce the cost and upgrade the quality of these critical vaccine components. This Phase IIB
resubmission focuses on commercial development of Scarab’s first vaccine product, the diphtheria mutant
toxin CRM197, an important component of highly successful conjugate vaccines, of significant interest to the
mission of the NIH. CRM197 has also shown promise as an anti-cancer agent. In high demand worldwide, it is
difficult to produce by batch technology, so supply is constrained and unreliable. For Scarab to enter the
vaccine market, cGMP-grade product is required. For maximum efficiency, Scarab proposes to outsource initial
production of cGMP-grade CRM197 to a company that will use Scarab’s innovative process in its GMP facility,
validating the process for GMP manufacturing. In a previous Phase 2 SBIR project, our process generated
high levels of pure recombinant CRM197 and yielded consistently high quality soluble and stable CRM197
protein. In particular, the A and B breakdown products are minor components of Scarab’s high-quality product.
The downstream process (DSP) from Phase 2 resulted in highly pure CRM197 now sold on the research
market. Since the original Ph2B proposal, new research has established (i) successful scale up to a 10L
productive fermentation process and (ii) a dramatically more modern and efficient DSP that is automatable, a
vital requirement for commercial viability.
Waisman Biomanufacturing (WB), a well-established not-for-profit cGMP contract development and
manufacturing organization, will perform production of cGMP-grade CRM197. WB was selected because it has
provided a very competitive quote, and has exceptional experience and expertise – it has manufactured (and
its clients have released) well over 300 clinical-grade products, with 5-10 investigational new drug applications
submitted by their partners each year. WB’s location in the same city as Scarab will facilitate technology
transfer and project oversight. The Specific Aims are:
1) Full development, scale-up and automation of the new DSP. Late-stage development of the fermentation
process at Scarab, including four pilot fermentations and downstream processing, technology transfer to WB,
and cGMP process development. 2) engineering GMP prototype run, specifications regarding purity and
potency established. 3) Three GMP production runs that formulize within-run and post-purification
specifications. 4) Final product validation that will include purity and potency assays (conjugation and human
immunogenicity). Stability studies by Nitto Avecia. Materials from GMP engineering and production runs will
provide potential customers with samples for evaluation. Project information will be used in a Biological Master
File at the FDA.

## Key facts

- **NIH application ID:** 10107730
- **Project number:** 5R44AI094823-06
- **Recipient organization:** SCARAB GENOMICS, LLC
- **Principal Investigator:** FREDERICK R BLATTNER
- **Activity code:** R44 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $1,000,000
- **Award type:** 5
- **Project period:** 2012-09-05 → 2023-02-28

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10107730

## Citation

> US National Institutes of Health, RePORTER application 10107730, Toxoid adjuvant CRM197 production in a stable reduced genome E. coli strain (5R44AI094823-06). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10107730. Licensed CC0.

---

*[NIH grants dataset](/datasets/nih-grants) · CC0 1.0*