How to choose a biological buffer for protein purification?

In recent years, protein chemistry and molecular biology have gradually developed, and many protein drugs have played an important role in clinical treatment, which has also become a hot spot in the development of the biomedical industry. Due to the high activity and low side effects of protein drugs, many diseases can be treated at the same time. The inherent advantage means that its purification is not so easy, because the protein must be kept inactive during the entire purification period. Therefore, the following factors must be taken into account in the preparation process.

1. PH value

The pH condition created in many experiments is 7.4. Purification is performed at this pH value. If instability occurs, the pH value needs to be changed so that it is soluble in solution but not degraded. When the pH of the solution is near the isoelectric point of the protein, because the protein surface has no net charge and is easy to aggregate, it will not be easily dissolved in the solution.

2. Buffer

First of all, it is necessary to ensure that there is a certain buffer capacity. In the case of the original pH value, the pKa value is also during this period. In addition, it is necessary to ensure that the concentration of the buffer solution can play the role of the buffer solution. Usually, the selected concentration is between 20 and 100 mM. However, it is important to note that none of the buffer systems used can affect protein activity. In addition, some buffers are susceptible to temperature changes, and changes in temperature lead to changes in the pH value. Therefore, when conducting experiments, it is necessary to consider whether the pH value is suitable for the experimental environment.

3. Salt ions

Some buffers contain sodium chloride, which is used to help stabilize the solubility of proteins. The concentration is generally 150mM. However, different protein purification steps actually require different salt ion concentrations.

4. Stabilizer

Stabilizers are added to the buffer, which are mainly used to help improve the solubility and stability during the purification process without interfering with the experiment. For example, glycerol is sometimes added to increase viscosity and help protein aggregation; sometimes a small amount of ions is added. Compounds, such as citric acid, are used to control ionic interactions and improve solubility.

Hubei New Desheng is a manufacturer specializing in the production and sales of biological buffers. At present, the buffer series products include TRIS, CAPS, BICINE, TRIS-HCL, TAPS, etc., with stable performance and high purity, which can effectively help improve efficiency. If customers have special customized needs, the company will fully cooperate to meet different needs. If you have biological buffer requirements, please click on the official website for more information!