# Process development for manufacturing eCD4-Ig

> **NIH NIH R44** · EMMUNE, INC · 2023 · $299,998

## Abstract

ABSTRACT
 eCD4-Ig is an antibody-like therapeutic for HIV that was constructed by fusing CD4 domains 1 and 2, and
an HIV coreceptor-mimetic sulfopeptide, to the N- and C-termini of the constant Fc region of an antibody.
eCD4-Ig will join an emerging class of long-acting therapeutics for treating and preventing HIV infection. As
a consequence of being built from parts of HIV’s receptors, eCD4-Ig neutralizes 100% of HIV isolates, and
even neutralizes simian immunodeficiency virus (SIV). Thus, eCD4-Ig presents fundamental barriers to
escape. This grant will fund the work that is needed in between cell line development and IND-enabling
studies to bring eCD4-Ig to the clinic. In Phase I, we will validate assays for critical quality attributes (CQAs)
including the sulfotyrosine content of the coreceptor-mimetic sulfopeptide, and then show that the eCD4-Ig
protein made by new cell lines meets pre-defined CQAs. In Phase II, we will optimize our upstream process,
downstream process, formulation buffers, and generate a GLP lot of eCD4-Ig protein for IND-enabling
studies. Features of the upstream process that will be optimized include media and feed selection, a
temperature shift, and fed-batch versus perfusion culture. Our downstream process is based on convective
chromatography, including for Protein A affinity chromatography, anion exchange (AEX) chromatography,
and hydrophobic interaction (HIC) chromatography. These processes efficiently remove poorly sulfated
eCD4-Ig, aggregates, and host cell protein (HCP). We will develop a formulation for intravenous injection,
and a high-concentration formulation with acceptable viscosity for subcutaneous injection. Production of a
GLP lot of protein for IND-enabling studies will be outsourced to a CMO after a technology transfer process.
These are the essential next steps towards bringing eCD4-Ig to the clinic.

## Key facts

- **NIH application ID:** 10603836
- **Project number:** 1R44AI170285-01A1
- **Recipient organization:** EMMUNE, INC
- **Principal Investigator:** MICHAEL DAVID ALPERT
- **Activity code:** R44 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2023
- **Award amount:** $299,998
- **Award type:** 1
- **Project period:** 2023-01-01 → 2023-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10603836

## Citation

> US National Institutes of Health, RePORTER application 10603836, Process development for manufacturing eCD4-Ig (1R44AI170285-01A1). Retrieved via AI Analytics 2026-05-26 from https://api.ai-analytics.org/grant/nih/10603836. Licensed CC0.

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