Expressing and purifying vaccine targets

Recombinant subunit protein vaccines are safe, potent, and well-characterized options that provide broad protection against multiple serotypes with low reactogenicity. Researchers working with recombinant protein-based subunit vaccines need to express their vaccine candidate proteins at high levels with a small, well-tolerated tag such as the 6xHis-tag to allow downstream purification. Choosing an efficient system for the optimal expression and efficient purification of the vaccine candidates is essential to speed up the vaccine development process.

Our BakPAK Baculovirus Expression System is ideal for obtaining high recombinant protein yields compared to other expression systems. It is also optimal for expressing proteins that are difficult to express in bacteria due to size, complexity, or post-translational modifications. BakPak technology is easy to scale up, has high protein expression levels, and is nonpathogenic to humans. The post-translational processing and folding of recombinant proteins produced in insect cells more closely resemble mammalian processes.

Our BacPAK Baculovirus Rapid Titer Kit uses a standard immunological assay to accurately determine baculovirus titers within 48 hours, while plaque and end-point dilution assays typically require 5–10 days, and is compatible with all commonly available (AcMNPV-based) baculovirus expression systems.

Once expressed from these purified constructs, his-tagged vaccine candidate proteins can be rapidly purified using our performance-validated cobalt and nickel resins or our revolutionary Capturem membrane-based purification technology. Capturem technology utilizes high-capacity nylon membranes immobilized with Ni-NTA, allowing rapid and easy his-tagged protein purification in under 30 minutes. In contrast, typical resin-based purification methods using immobilized metal affinity chromatography (IMAC) columns require several hours to complete due to long column equilibration/binding times and slow diffusion of large macromolecules through the resin bed.

Learn about the many ways Capturem technology can help speed up your research.

Benefits

• Fast, convenient workflow—5- to 30-min room-temperature protocol using convenient spin columns/plates/filtration devices featuring Capturem nickel-functionalized membranes
• High purity—small bed volumes trap fewer contaminants
• Compatible with a wide range of additives—including EDTA, DTT, BME, glycerol, TCEP, etc. (see compatibility table)
• Versatile—easy purification from mammalian/bacterial cell lysates and supernatants

Numerous laboratories have already used our TALON resin and Capturem technology for their vaccine development research:

Title	Link	Product
Protein purification
SARS-CoV-2
A rapid, point of care red blood cell agglutination assay for detecting antibodies against SARS-CoV-2		Capturem His-Tagged Purification Miniprep Kit
Dynamics of neutralizing antibody titers in the months after SARS-CoV-2 infection		TALON resin
Functional SARS-CoV-2-specific immune memory persists after mild COVID-19
Pre-existing and de novo humoral immunity to SARS-CoV-2 in humans
SARS-CoV-2 and bat RaTG13 spike glycoprotein structures inform on virus evolution and furin-cleavage effects
Ultra-sensitive high-resolution profiling of anti-SARS-CoV-2 antibodies for detecting early seroconversion in COVID-19 patients
Other viruses
Recombinant adenovirus carrying a core neutralizing epitope of porcine epidemic diarrhea virus		Capturem His-Tagged Purification Miniprep Kit
Generation of the recombinant antigen LC3 from Entamoeba histolytica linked to mutated exotoxin A (PEΔIII)