Application of HEPES in biological science

攝影師:Martin Lopez:
Article Author: YJ Chen / R&D Mannager of Fine Chemical Division

What is HEPES Biological Buffer?

HEPES biological buffer full chemical name is N-(2-Hydroxy ethyl)-Piperazine ethane Sulfonic acid (hereinafter referred to as “HEPES”), its function is mainly to buffer the surrounding pH value changes caused by the environment and biological metabolic waste etc,. so as to maintain the biological environment at an appropriate pH value. HEPES buffer ranges from pH 6.8 to pH 8.2. At present, it is widely used in cell experiments, bacterial culture, detection reagents, biological preservation liquids and other products.

HEPES and cell culture

In the 3D cell culture patent (CN113372580A, 2021), HEPES is added to remove calcium ions and maintain the pH of the medium, allowing cells to grow in a range that is relatively unaffected by external influences. Some patents also mention that HEPES is added to wound dressings or artificial skins (CN113209365B, 2022), so that the medicines in the dressing and the wound can be maintained in a stable state, avoiding deterioration due to tissue fluid or storage reasons. It is also commonly used to replace KCl and NaCl.

It is mentioned in the patent (WO2021168764A1, 2021) that adding HEPES to the mitochondrial cryopreservation agent can effectively protect the mitochondrial structure and maintain its function after thawing, and provide the effect of maintaining pH in the Isotonic solution.


HEPES use for animal husbandry

In animal husbandry, selecting the required genes and performing artificial insemination is a very important link, but after the sperm and eggs of the animals are taken out from the farm, there will be preservation problems in the process of delivering them to the laboratory.

Experiments have confirmed that adding an appropriate concentration of HEPES (10mM) in the transport environment can prolong the survival time of sperm and eggs in a fixed environment and help maintain the survival rate of sperm and egg cells arriving in the laboratory (CN112980778B, 2021). Artificial breeding can greatly improve the probability of fertilization, and maintain genetically screened animal breeds suitable for breeding.


Clinical trial use of HEPES

In clinical practice, whole blood preservation is very important, and fresh whole blood samples can provide a variety of clinical applications, such as cytokines, immunoassays, stem cells isolated from umbilical cord blood to peripheral blood to provide transplantation(EP3177924B1, 2020), etc.

It is mentioned in this patent that after extraction of human blood, the formulation of whole blood sample stabilizer is added with high concentration of HEPES (48mM) to prevent large changes in pH value and prevent the activation of neutrophils such as neutrophils, sudden oxidation and cell necrosis and apoptosis.

Further Reading: Why use HEPES?
Further Reading:HEPES handling and storage tips that you must know

Release date:2022.05.27