Separation & Purification of Viruses by Ion Chromatography

Alfa Chemistry provides you with the purification and separation of viruses and their vectors. You can deliver the project to us at any stage of the experiment. Compared with the macroscopic world, the structure of the virus is much more complicated, so its separation and purification are also more difficult. With a series of reagents and equipment for virus separation and purification, we are ready to serve you.

Introduction

Although viruses are microorganisms, their size is larger than traditional protein and nucleic acid biomolecules, generally between 20-100 nanometers. People need to use an electron microscope to see the structure and appearance of the virus. The emergence of genetic drugs and good therapeutic effects have led to the continuous deepening of drug research using viruses as carriers. For example, adeno-associated virus (AAV), as a linear single-stranded DNA virus, is mainly used to treat genetic diseases such as Parkinson's disease, muscular dystrophy, and hemophilia B. The UniQure company used AAV1 as a vector, carrying the LPL gene, for the treatment of severe or recurrent pancreatitis caused by lipoprotein lipase deficiency. The residual protein, nuclease, or nucleic acid in the virus will not only affect the transfection effect, but also cause an immune response, so it needs to be separated and purified. The separation and purification of viruses mainly include ultracentrifugation, chromatography, chemical reagents and ultrafiltration.

Ion exchange chromatography

The isoelectric point of the virus as a special pH value is determined by the sum of the amino acid side chains exposed on the surface of the capsid and the static charge of the N-terminal or C-terminal packaged in the carrier. The level of the isoelectric point determines the type of ion chromatography used. When the pH of the solution is greater than the isoelectric point of AAV, anion chromatography is used for separation. Under a certain concentration of NaCl gradient elution condition, the viral vector containing certain gene passing through an anion column can obtain an absorption peak with better resolution, and the recovery rate can reach 99%. When the pH of the solution is less than the isoelectric point of AAV, cation chromatography can be used for separation. In addition, hybrid chromatography can be used for the viruses which are difficult to combine with ion exchange chromatography, or even if combined, it is difficult to establish suitable conditions for elution. For example, a combination of ceramic hydroxyapatite chromatography and cation exchange column.

Advantages

• The operation process is flexible and can be adjusted according to actual needs, which can purify large-volume cell lysates or crude virus extracts on a large scale.
• The purification process helps maintain the biological activity of the virus.
• All operations can be completed in a closed system with less pollution.
• It has a wide range of applications and is suitable for a variety of viruses.

The virus isolation and purification service provided by Alfa Chemistry allows you to obtain safe, effective and inexpensive virus vectors to better meet the needs of laboratories, clinical trials and market. Our customers can directly contact our experts and provide timely feedback on any online inquiries. If you are interested in our services, please contact us.