Composition of Shelf-stable Plasmid DNA/PEI Particles with Defined Sizes for Virus Production and Method for Preparation of the Same

Case ID:
C16592

Unmet Need

Gene therapy has become increasingly popular for treating genetic and acquired diseases, with nearly 1000 ongoing clinical trials globally. These therapies are typically delivered via viral vectors, which are produced in vector production cell lines following transient transfection using nanoparticles such as those made of plasmid DNAs (pDNA) and polyethyleneimine (PEI), a polycation. With an increasing number of clinical trials and studies, the production demand of viral vectors has increased substantially. However, the current standard processes for preparing cell transfection nanoparticles require batch-mode production immediately before cell transfection for vector manufacturing. This pDNA/polycation nanoparticle preparation process is prone to lower consistency at the industry manufacturing scale, resulting in higher variability in production yield of viral vectors, comparing to the bench tests. There is a need for a low-variability, high transfection efficiency, and off-the-shelf pDNA/polycation nanoparticles that can enable large scale manufacturing of viral vectors.

 

Technology Overview

Johns Hopkins researchers have developed a scalable method of producing shelf-stable pDNA/polycation particles in the range of 50 to 1000 nanometers. They have also determined the optimal composition and size of the pDNA/polycation particles for maximum in vitro transfection efficiency, with an optimal size of 400 to 500 nanometers. The off-the-shelf formulation of the 400-nm pDNA/PEI particles demonstrated superior shelf stability at -80 degrees Celsius with preserved physical properties and transfection activity. The particle formulation was validated using lentiviral vectors and demonstrated consistent yield equivalent to standard manual preparation methods, drastically streamlining the viral manufacturing process and enabling reliable large-scale production of lentiviral viral vectors.

 

Stage of Development

Method development completed.

Patent

Provisional filed.

Publication

Hu Y, Zhu Y, Sutherland ND, Wilson DR, Pang M, Liu E, Staub JR, Berlinicke CA, Zack DJ, Green JJ, Reddy SK, Mao HQ. Size-Controlled and Shelf-Stable DNA Particles for Production of Lentiviral Vectors. Nano Lett. 2021 Jul 14;21(13):5697-5705.

Patent Information:
Title App Type Country Serial No. Patent No. File Date Issued Date Expire Date Patent Status
COMPOSITION OF SHELF-STABLE PLASMID DNA/POLYETHYLENEIMINE (PEI) PARTICLES WITH DEFINED SIZES FOR VIRAL VECTOR PRODUCTION AND METHOD FOR PREPARATION OF THE SAME PCT: Patent Cooperation Treaty European Patent Office 22756830.0   2/16/2022     Pending
COMPOSITION OF SHELF-STABLE PLASMID DNA/POLYETHYLENEIMINE (PEI) PARTICLES WITH DEFINED SIZES FOR VIRAL VECTOR PRODUCTION AND METHOD FOR PREPARATION OF THE SAME PCT: Patent Cooperation Treaty Japan 2023-549082   2/16/2022     Pending
COMPOSITION OF SHELF-STABLE PLASMID DNA/POLYETHYLENEIMINE (PEI) PARTICLES WITH DEFINED SIZES FOR VIRAL VECTOR PRODUCTION AND METHOD FOR PREPARATION OF THE SAME PCT: Patent Cooperation Treaty United States 18/546,222   8/11/2023     Pending
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For Information, Contact:
Lisa Schwier
lschwie2@jhu.edu
410-614-0300
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