How does recombinant glycerol kinase cope with detection interference?

Triglycerides (TG) are an important component of blood lipids, and their concentration levels are key indicators for assessing the risk of cardiovascular disease. For many years, clinical detection of triglycerides has mainly relied on enzyme-linked immunosorbent assay (ELISA) kits. One of the core enzymes in this method, glycerol kinase (GK), is responsible for catalyzing the generation of glycerol-3-phosphate from glycerol, initiating a series of enzymatic reactions, and ultimately determining TG content through photometry. However, traditional natural sources of glycerol kinase are often accompanied by interfering enzymes such as NADH oxidase and catalase, which can consume reaction substrates or produce side reactions, resulting in large deviations and poor reproducibility of detection results. Faced with the pain points in this industry, recombinant glycerol kinase has emerged, leading clinical testing towards a new era of precision with its high purity and low interference characteristics.

1, Interference enzyme problem: the "invisible killer" of traditional detection

In the process of enzyme-linked immunosorbent assay for detecting triglycerides, glycerol kinase needs to work synergistically with glycerophosphate oxidase, peroxidase, and other enzymes to produce chromogenic substances through the Trinder reaction. However, if the glycerol kinase preparation contains NADH oxidase, it will non specifically degrade NADH (reduced coenzyme I), resulting in a decrease in absorbance; If catalase is present, it will decompose the hydrogen peroxide generated in the reaction, reducing the color intensity. The presence of these two interfering enzymes can cause deviation from the standard curve in mild cases and completely invalidate the detection results in severe cases. Natural sources of glycerol kinases (such as those extracted from bacteria or fungi) are often difficult to completely remove due to their complex extraction processes and purification difficulties, making them a bottleneck that limits detection accuracy.

2, Recombinant technology: purifying from the source and creating 'pure enzymes'

The breakthrough in recombinant DNA technology has brought revolutionary changes to the production of glycerol kinases. By introducing the glycerol kinase gene into engineering bacteria such as Escherichia coli and optimizing expression conditions, recombinant glycerol kinase can achieve efficient and controllable expression. More importantly, the recombinant production system avoids the coexistence of multiple enzymes in natural strains, and subsequent precision purification processes such as chromatography and ultrafiltration can effectively remove interfering components such as NADH oxidase and catalase. The purity of recombinant glycerol kinase is over 95%, which is much higher than that of natural enzyme preparations, cutting off the pathway of interfering enzyme contamination from the source.

3, Stability Enhancement: Scientific Response to Temperature Sensitivity Issues

Although recombinant glycerol kinase has significant advantages in purity, its stability in environments above 45 ℃ is poor, which was once one of the obstacles to its widespread application. Through protein engineering modifications such as site directed mutagenesis and rational design, researchers have successfully improved the thermal stability of enzymes. At the same time, optimizing the buffer system and protective agent combination in the formulation (paying attention to avoiding the negative effects of trehalose) further extended the storage time and usage duration of enzyme activity. Nowadays, high-quality recombinant glycerol kinase can maintain activity for a long time at 4 ℃ and perform robustly under room temperature detection conditions, fully meeting the storage and operation requirements of the reagent kit.

Hubei Xindesheng Material Technology Co., Ltd. has always adhered to the production and operation philosophy of "quality first, technology leadership" and "innovation and pursuit of excellence", continuously researching and developing reagents needed in the diagnostic field. Since its establishment in 2005, it has a history of nearly 20 years and is deeply trusted and supported by users. The diagnostic enzyme reagents independently developed and produced by Desheng have been sold to various parts of the world. If you have any related procurement needs in the near future, please feel free to contact us!