What you should know about the core biological material of IVD - enzymes

With the leapfrog progress of the in vitro diagnostic reagent industry, a complete industrial chain has now been formed, including the supply of raw materials from upstream manufacturers, the research and development, production and sales of midstream reagent products, and downstream market demand. The supply quality of core raw materials from upstream manufacturers is crucial, directly determining the development of finished reagent products. For example, upstream core raw materials such as enzymes, antigen antibodies, buffering agents, blood vessel additives, chromogenic substrates, luminescent reagents, etc. must all require refined production. Below, we will introduce to you the widely used core raw material - enzymes.

1、 Basic principles for the research and development of raw material enzymes

The main way to obtain enzymes is through purification, so protein purification technology and antibody preparation technology have become the key and fundamental principles for the development of active raw materials, which may involve multiple disciplines.

2、 Scale preparation of raw enzyme

The entire preparation process includes several steps, including preparing raw materials, pre-treatment, initial purification, and precise purification. The preparation of raw materials is directly obtained by selecting cells from animals and plants or through genetic means. The pretreatment of raw materials is to use some means to break the cell tissue, or repeatedly freeze and thaw or treat with surfactant. However, it is necessary to pay attention to the strength of breaking histiocyte. Sometimes the strength is small, it will not be fully released, and the strength is large, which will make it variable. The raw materials will gradually become turbid after breaking, which can be removed by centrifugation.

The initial purification is carried out through precipitation or centrifugal filtration, commonly used methods include ammonium sulfate precipitation or the use of some organic solvents for precipitation. Precise purification methods mainly include various separation chromatography based on molecular structure size, distribution or hydrophobic properties, including ion exchange chromatography, hydrophobic chromatography, etc.

3、 Preservation of raw enzyme

The factors that affect the solution state of enzyme active raw materials generally include the change of PH value, ionic strength, storage conditions, repeated freezing and thawing times, etc. Generally, the stability can be maintained by adding a series of buffer systems, inhibitors or reducing agents, and it is very common to freeze dry storage.

4、 Quality control of raw enzyme

The key to controlling enzyme quality is to identify the raw enzyme after batch purification, including concentration and purity, activity identification. The commonly used methods are enzyme-linked immunosorbent assay or protein quantification. The enzyme raw materials sold from the factory should be of high purity, high activity, and meet the standards for both appearance and concentration.