Plasmid DNA Manufacturing

PLASMID MANUFACTURING SERVICES

Xpress Biologics scientists combine over 30 years of experience in the field of gene therapy and have implemented a cost effective and flexible plasmid DNA manufacturing platform with dedicated QC tests for R&D, preclinical validation as well as early phase clinical trials. Our production scale ranges from 20 mg up to 30 g of supercoiled (SC) plasmid with a 95 to 98 % purity independently of the plasmid grade selected.

Our plasmid manufacturing platform encompasses:

  • High cell density E. coli fermentation with production yields up to 1.8 g/L of plasmid
  • Optimized & scalable alkaline lysis process allowing high SC content, low gDNA (< 5.0 %) and RNA (< 5.0 %) content.
  • Chromatographic purification step for effcient open ciruclar form, endotoxine and residual RNA, gDNA removal
  • “off-the-shelf” rapid method for R&D or High-Quality (HQ) grade production

R&D-Grade Plasmid DNA

Xpress Biologics offers a flexible platform providing high quality R&D grade DNA (>95% SC, endotoxin≤ 20 EU / mg DNA) with cost effective pricing and competitive delivery time from 20 mg up to 2g of purified material.
R&D grade plasmid can be used as Drug substance/DNA vaccine or as viral vector manufacturing starting material (e.g., AAV, adenovirus, or lentivirus) for in vitro or in vivo applications.

Deliverables

  • Glycerol Cell Bank
  • Purified product
  • Certificate of analysis

High Quality-Grade Plasmid DNA

Xpress Biologics can provide HQ grade plasmid for preclinical validation as well as early phase clinical trials. HQ grade plasmid may be used as critical starting material for the production of  GMP grade viral vectors as well as Drug substance/DNA vaccine for in vitro or in vivo applications.

Deliverables:

  • Characterized Research Cell Bank (RCB)
  • Purified product
  • Certificate of analysis
  • Batch record including production method, raw materials list, production check-lists and TSE/BSE Statement
  • QA oversight

GMP-Grade Plasmid DNA (on line Q2 2022)

Xpress Biologics will provide GMP-grade plasmid DNA in segregated ISO-classified manufacturing laboratories .

The targeted scale will range from 10 to 100 liters yielding up  to 60 g of highly pure supercoiled plasmid DNA. Our GMP grade plasmid may be used as drug substance, DNA vaccine or starting material for viral vector manufacturing for all clinical phases and for commercial applications

Deliverables:

  • GMP E. coli Working and Master Cell Bank
  • Scalable GMP plasmid manufacturing process
  • Purified product
  • Certificate of analysis
  • Batch record including production method, raw materials list, production check-lists and TSE/BSE Statement
  • QA oversight
  • QC and Release by Qualified Person (QP)

 

Standard plasmid quality controls

Tests Methods
Physcal state pH, conductivity and appearance
Identity Restriction mapping
DNA sequencing
Content Spectrophotometry - A260
Purity Spectrophotometry - ratio A260/A280
Topology by agarose gel electrophoresis (SC, OC, CC)
Topology by IEx-HPLC (SC, OC)
Impurity Residual host cell proteins by µBCA
Residual host cell DNA by q-PCR
Residual host cell RNA by agarose gel electrophoresis or HPLC
Safety Endotoxin content by LAL
Bioburden
Potency Specific test

 

Plasmid grade comparison

Plamsid manufacturing services R&D HQ GMP (2022)

Screening for optimal growth conditions

(for non standard plasmids/strains)

x

 

 
Shake flask production x    
High-productivity fermentation production (up to 1g/L) x x x
Optimized alkaline lysis for high SC content (>95%) x x x
Chromatographic purification x x x
Platform manufacturing process x x x
Certificate of analysis (CoA) x x x
E. coli glycerol cell bank generation x x x
Manufacturing summary report x x x
E. coli research cell bank generation   x x

Manufacturing batch record

  x x

RNAse free and TSE/BSE certificate

  x x

Change control management

  x x

QA oversight

  x x

QA release+

  x x

E. coli master cell bank generation

    x

GMP production suites 

    x

Environmental monitoring

    x

Classified production suites

    x

Master batch records

    x

QMS following GMP guidelines

    x

QA release

    x