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Veterinary Antigen Purification
Antigen purification is the process of separating and purifying the antigen from biological tissues or the mixture generated for synthesizing antigens, with the aim of removing unrelated components and obtaining a highly pure antigen. BioVenic offers various purification methods for different types of veterinary antigens, including proteins, peptides, nucleic acids, and inactivated pathogens like viruses and bacteria, etc.
Our Services
Cell Lysis
For a variety of veterinary antigens, such as recombinant proteins, nucleic acids, bacterial polysaccharides, and viruses, cell lysis is required to release the antigens from cells, facilitating subsequent purification steps. BioVenic offers various methods for cell lysis, allowing you to choose the one that best suits your cell type while ensuring minimal denaturation and degradation of the antigen.
Mechanical Disruption
Chemical Lysis
Ultrasonic Disruption
Chromatography
BioVenic offers antigen purification services utilizing chromatography technology. Chromatography achieves the separation and purification of biological molecules by leveraging the interactions between the target molecule and a stationary phase (typically resin or gel) and a mobile phase (usually liquid). Through this process, we ensure the acquisition of high-purity and high-quality antigen samples, suitable for further development and production of veterinary vaccines.
| Services | Description |
|---|---|
| Gel Filtration Chromatography |
Antigen samples pass through a porous agarose gel with defined pore sizes, separating components based on differences in molecular size. Common gels used include calcium phosphate gels, zinc hydroxide gels, and magnesium pyrophosphate gels. This method ensures the removal of free nucleotides and truncated transcription products, especially when dealing with mRNA prepared through in vitro transcription. |
| Ion Exchange Chromatography |
Suitable for the separation and purification of all charged antigens. By adjusting the pH to certain conditions, it is possible to separate viral particles based on their different charge properties. |
| Affinity Chromatography |
leveraging the specific affinity between the antigen and a fixed ligand or antibody for separation. It is an ideal choice for high-purity and antigen-specific purification of large molecules. |
| Size-Exclusion Chromatography | Separates proteins based on their molecular size, allowing the separation of larger proteins while removing smaller proteins and contaminants. |
| Reverse-Phase Chromatography | Employs hydrophobic interactions to separate proteins or peptides with varying hydrophobicity, facilitating the purification of hydrophobic proteins or peptides. |
Fig.1 How gel filtration chromatography works. 1
Precipitation
BioVenic employs the simple and straightforward precipitation method, utilizing specific precipitants that interact with the target antigen, causing the antigen to precipitate while impurities remain in the supernatant. Through centrifugation and other procedures, a precipitate containing high-purity antigen is separated.
| Services | Description |
|---|---|
| Inorganic Salts |
Ammonium sulfate is a commonly used salt in antigen purification as it reduces the solubility of proteins in the solution. For mRNA obtained through in vitro transcription, lithium chloride precipitation is employed to remove the majority of unbound nucleotides and enzymes. BioVenic meticulously controls parameters like temperature, pH, and antigen concentration to prevent excessive salting-out. |
| Organic Solvents | The formation of multimers between virus particles and polyethylene glycol is utilized to precipitate the virus, subsequent centrifugation achieves this. |
Centrifugation
BioVenic utilizes the principle of centrifugation to effectively separate and precipitate antigens to the bottom. Through targeted sampling or separation of the antigen precipitate at the bottom, high-concentration and high-purity antigen samples will be obtained. For viral antigens, their size and density differ from those of cells, biological macromolecules (such as DNA, RNA, and proteins), and other host components in contaminated virus samples.
| Services | Description |
|---|---|
| Density Gradient Centrifugation |
Used to distribute virus particles into equally dense layers of a density gradient medium. The desired particles are then withdrawn from the medium layer, achieving the goal of separation and purification. Common gradient media include cesium chloride and sucrose. |
| Differential Centrifugation |
Exploits the varying sedimentation rates of particles with different sizes and densities, effectively removing impurities like host cell debris. Useful for rapidly processing large sample volumes and aids in the separation of cellular organelles and proteins from cell lysates, ultimately resulting in antigen precipitation. |
Protein Labeling
Protein labeling is a widely used method in protein purification, involving the attachment of short peptides to the target protein. These labels enhance the solubility and stability of recombinant proteins, facilitating their purification and identification. BioVenic provides antigen purification services utilizing this method, enhancing the precision and efficiency of the purification process.
| His-Tag (Histidine Tag) | GST-Tag (Glutathione S-Transferase Tag) | FLAG-Tag |
Solid-Phase Extraction
Solid-phase extraction (SPE) combines the advantages of liquid-solid extraction and chromatography. It is used by BioVenic for the enrichment and purification of proteins, nucleic acids (DNA and RNA), and peptides, where a solid phase (adsorbent or resin) binds analytes or impurities through strong yet reversible interactions, allowing the reliable and rapid extraction of antigens from complex samples.
Why Choose Us?
A variety of purification platforms available for different antigen components.
High purity and high yield.
One-stop service offering all antigen-related services throughout veterinary vaccine development process.
Antigen purification is necessary to support the subsequent veterinary vaccine development, with significant impacts on the immunogenicity and safety of the final product. BioVenic has established diverse antigen purification platforms to meet specific research and develop needs. If you require antigen purification in veterinary vaccine development, please contact us and we will leverage our extensive experience to provide you with satisfactory services!
Reference
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From Elsevier:
https://ars.els-cdn.com/content/image/3-s2.0-B9780123847300001099-f00109-07-9780123847300.jpg