Fermentation for Oligonucleotides and Nucleic Acids

As a leading CDMO, BOC Sciences provides one-stop services to manufacture oligonucleotides and nucleic acids (such as plasmid DNA). with the large scale fermentation capabilities and advanced downstream processes, we provides one-stop fermentation services to meet our customers' commercial production needs.

Learn more about Fermentation Products.

Introduction of Oligonucleotides and Nucleic Acids

Oligonucleotides are short-stranded nucleic acid (DNA or RNA) molecules that are usually synthesized in the laboratory by solid-phase chemistry. Early innovations in biotechnology, such as fermentation, were key stepping stones to the birth of oligonucleotide synthesis. Nucleic acid metabolism is the process of DNA and RNA synthesis and degradation. Nucleic acids are polymers of nucleotides. The development of recombinant and bioengineering technologies has made possible the production of nucleic acid-related molecules through fermentation processes.

Currently, the rapid development of mRNA technology has led to a high level of interest in the field of nucleic acid drugs. Nucleic acid drugs include antisense oligonucleotides (ASO), small interfering RNA (siRNA), micro RNA (miRNA), small activating RNA (saRNA), messenger RNA (mRNA), aptamer, etc. To date, the novel coronavirus disease is still spreading rapidly worldwide and vaccines are an important way to control the spread of this virus, of which the development of nucleic acid vaccines is a hot topic. Nucleic acid vaccines generally include viral vector vaccines, plasmid DNA vaccines and mRNA vaccines, which have advantages such as time cost and production scale compared to traditional vaccines.

Fermentation Production of Plasmid DNA

A plasmid is a small extrachromosomal DNA molecule within a cell that can replicate independently. Plasmid DNA preparation is based on the sequence design of the transcriptional template, and the preparation method usually employs plasmid DNA amplification. Plasmid amplification is achieved by fermentation production using engineered E. coli as the host. Therefore, plasmid DNA production starts with the establishment of an engineered bacterial seed bank.

Fermentation for Oligonucleotides and Nucleic Acids

The major steps in the production of plasmid DNA:

Fermentation for Oligonucleotides and Nucleic Acids

Strain Culture

Starting with resuscitation of frozen strain, the strain is cultured through multiple stages of amplification to achieve the required number of strains for production scale culture.

Fermentation

Strain fermenting at the desired production scale achieves sufficient replication of plasmid DNA to meet the final plasmid yield requirements.

Harvest

Since a large number of replicated plasmids are present in the bacteria, collection and lysis of bacteria is required to release plasmid DNA for subsequent isolation and purification. Continuous flow centrifuges are commonly used in industry to remove bacteriophage debris and some flocs to obtain a clarified harvest solution.

Purification

The refinement and purification process is performed by tangential flow filtration for concentration followed by chromatographic purification.

Gel chromatography removes impurities based on molecular size.
Affinity chromatography selectively adsorbs biomolecules of interest.
Anion exchange chromatography removes trace impurities such as bacterial endotoxins.

Our Services

Our Advantages

One-stop fermentation service from laboratory scale, pilot scale to large scale
Efficient upstream and downstream process development
Large-scale industrial fermentation tanks of different volumes from 4,000L to 12,000L, over 100,000 liters capability in total
Well established quality system, Quality-by-Design (QbD) and process analytical techniques (PAT)
Strict aseptic verification procedures throughout the fermentation process
GMP certified fermentation production
Investment in environmental protection measures and skills