# Flexible Platform for End-to-end Manufacturing of Gene Therapies to Advance Development of Treatments for Ultra-rare Diseases

> **NIH FDA R01** · MASSACHUSETTS INSTITUTE OF TECHNOLOGY · 2020 · $400,000

## Abstract

SUMMARY / ABSTRACT
The success of gene therapies for addressing inherited disorders has been demonstrated by FDA
approval of AAV-based LUXTURNA® and ZOLGENSMA®. There is great hope that gene
therapies will also be able to address more of the 10,000+ monogenic diseases that have been
identified to date. Gene therapy developers face challenges in cost and access to manufacturing
capacity to produce the gene therapy product. The development of treatments for ultra-rare
genetic diseases, which occur in fewer than one person in a million, are especially disadvantaged
as these therapies are not typically of commercial interest to biopharmaceutical manufacturers.
There is an urgent need for innovation in meeting the need for developers of gene therapies for
ultra-rare diseases to gain access to GMP manufacturing of small doses of drug product. We
propose a flexible, cost-efficient, end-to-end manufacturing platform for production of a small
number of doses of AAV gene therapy products. We use the method of triple transfection in HEK
cells, which can be adapted to multiple genetic diseases by switching the disease-specific
genomic insert in one of the plasmids, as well as adapted to multiple AAV serotypes by switching
the AAV-specific plasmids. Our easy-to-use microbioreactor platform grows cells to high-density
in perfusion. We will optimize the the triple-transfection process for high-density cell cultures to
increase bioreactor productivity and generate material for treating small numbers of patients.
Additionally, our platform addresses the need for innovation in downstream purification of AAV
products. Empty capsids lacking the gene of interest can compose 10% - 90% of AAV produced
in cell cultures. Current vector purification approaches require specialized equipment which may
not be readily available in the standard laboratory. We will develop a novel downstream
purification method based on electrokinetic separation which greatly simplifies elimination of
empty AAV capsids. Finally, as the supply of GMP-grade plasmid needed for the triple transfection
process can also be a bottleneck, as well as significant expense, we will demonstrate that our
platform is capable of incorporating plasmid manufacturing, including production and purification,
to provide an end-to-end solution to generation of AAV therapies. This platform, will (1) expedite
the availability of AAV treatments for ultra-rare diseases by increasing the volumetric yields of
upstream process with minimization of the cost; (2) reduce the time, cost and labor associated
with the downstream AAV purification; and (3) incorporate plasmid production to ensure timely
and low-cost supply of this critical raw material. This platform will democratize access to viral
vector for developers of gene therapies for ultra-rare diseases.

## Key facts

- **NIH application ID:** 10201300
- **Project number:** 1R01FD007226-01
- **Recipient organization:** MASSACHUSETTS INSTITUTE OF TECHNOLOGY
- **Principal Investigator:** JONGYOON HAN
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** FDA
- **Fiscal year:** 2020
- **Award amount:** $400,000
- **Award type:** 1
- **Project period:** 2020-09-10 → 2023-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10201300

## Citation

> US National Institutes of Health, RePORTER application 10201300, Flexible Platform for End-to-end Manufacturing of Gene Therapies to Advance Development of Treatments for Ultra-rare Diseases (1R01FD007226-01). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10201300. Licensed CC0.

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