Vaccine Manufacturing with SUMO

Vaccine Manufacturing as SUMO Fusions

Success of mRNA vaccines has taken the world by a storm. However, many vaccines, especially animal vaccines, have not adopted mRNA approaches. SUMO mediated dramatic enhancement of protein production in E. coli provides a rapid and inexpensive approach to develop protein vaccines, especially for the veterinary markets.

The SUMO Tag

SUMO (Small Ubiquitin-like Modifier) is a member of the ubiquitin family, composed of a flexible N-terminal region followed by a ball-shaped ubiquitin-like fold. SUMO’s hydrophobic core improves correct folding of the fused polypeptides, and its hydrophilic surface keeps the partner protein soluble. SUMO protease recognizes SUMO structure to cleaves at the junction to generate desired N-terminus of the polypeptide.

Figure 1: The unique string and ball structure of SUMO allows for improved expression due to a hydrophilic outer surface and a hydrophobic core.

Understanding SUMO

It is important to understand why the SUMO system has proven to be the best among all other fusion systems (consult LifeSensors’ SUMO Search Engine). The chaperoning properties of SUMO promote correct folding and protect the vaccine from degradation. These factors contribute to dramatically enhance protein production and improve quality of the vaccine. E. coli provides an attractive platform to express animal vaccines for its low cost. The SUMO system provides facilities to increase protein yield and is a tool to rapidly purify the antigen. LifeSensors can guide scientists about the most suitable SUMO tag and the SUMO protease to efficiently remove SUMO to reduce cost of goods. LifeSensors can guide you from your cloning strategy to selection of strains/cell lines for the best expression and establish a non-GMP process from gene to vaccine. The process can be easily adapted for downstream GMP production of vaccine.

Figure 2: The Principle of the system. SUMO Protease cleaves the SUMO 6xHis tag to separate the expressed vaccine to generate native vaccine. In many cases SUMO tag is not removed and SUMO-vaccine is as efficacious. SUMO tag does not influence immunogenicity of the fused protein. The 6xHis is used to assist in purification while SUMO significantly improves expression.

LifeSensors pioneered the application of SUMO system to enhance production of polypeptides as published by Marblestone et. Al, Protein Sci. 2006. SUMO system is superior to several other tags: 6xHis, Ubiquitin, MBP, GST, and TRX. More recent studies [1],[2],[3],[4],[5] have shown a range of applications through expression of antimicrobial, cytotoxic peptides, and anti-thrombotic peptides.

Removal of the polypeptide from the tag is costly or impossible due to the inability of the proteases to cleave and provide the desired amino acid sequence. LifeSensors’ SUMO proteases recognize the SUMO structure and does not discriminate between fused polypeptide cleavage sites by generating the desired N-terminal sequence. As shown in the table below (Table 1), the cost of enzymatic cleavage of polypeptide is a major factor that has been a hurdle in adopting recombinant methods for polypeptide production. LifeSensors has developed the most efficient SUMO protease to reduce the cost of goods. As shown in the table below, SUMO mediated manufacturing of the polypeptide is the most cost-effective method. We encourage inquiries to establish cost-effective pilot scale manufacturing of polypeptides. The manufacturing process can be seamlessly transferred to GMP facilities.

Table 1: Several vaccines have been previously expressed using the SUMO system. SUMO is a powerful expression mechanism to increase yields and improve solubility. References are included at the end of the page.

SUMO is a highly conserved eukaryotic protein. In LifeSensors’ experience SUMO tag is not immunogenic when injected into rabbits. Therefore, it is quite feasible to inject SUMO-fusion vaccine as an immunogen, especially for veterinary applications. This simplifies the manufacturing process and reduces the cost of vaccine production. LifeSensors has established a curtailed list that allows you to search a variety of papers that describe the application of SUMO to express proteins, polypeptides and nanobodies, see below.

To use, simply utilize the drop down arrows in order to search for your target of interest or application category. Lifesensors SUMO technology has been utilized for the expression of many different targets and continues to be utilized worldwide.

AB7002H: HRP-labeled Anti-SUMO/SUMOstar Antibody

AB-7002-0051

Antibodies, SUMO Antibodies, SUMO Protein Expression Systems

pE1001K: pE-SUMOpro Kan Vector

PE-1001-K020

Bacteria, SUMO Protein Expression Systems

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Vaccine Manufacturing with SUMO

Vaccine Manufacturing as SUMO Fusions

The SUMO Tag

Understanding SUMO

AB-7002-0051

Antibodies, SUMO Antibodies, SUMO Protein Expression Systems

PE-1001-K020

Bacteria, SUMO Protein Expression Systems

PE-1001-A020

Bacteria, SUMO Protein Expression Systems

AB-0701-0250

DUB and Ubiquitin Like Isopeptidase Antibodies, SUMO Antibodies, SUMO Protein Expression Systems

PE-1106-0020

Bacteria, SUMO Protease Enzymes, SUMO Protein Expression Systems, SUMOstar Protease

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