Nuclease Activity Detection Kits

Deoxyribonucleases (DNases) and ribonucleases (RNases) are hydrolytic enzymes that degrade DNA and RNA by cleaving phosphodiester bonds. They are widely present in laboratory environments, raw materials, and consumables, and may also persist as process-related residues during biopharmaceutical manufacturing. Residual nucleases can compromise nucleic acid integrity, interfere with analytical workflows, and pose potential safety risks due to immunogenicity or unintended nucleic acid degradation. Therefore, sensitive and reliable detection of residual nuclease activity is critical throughout biopharmaceutical research, development, and production.

Conventional methods such as gel electrophoresis and UV spectrophotometry are commonly used to detect residual DNase and RNase activity, but they have limitations in sensitivity, quantification accuracy, throughput, or instrumentation requirements. In contrast, fluorescence probe–based assays offer high sensitivity, rapid detection, and precise quantitative measurement, making them a preferred approach for reliable residual nuclease activity detection.

Following rigorous methodological validation, ACROBiosystems has independently developed fluorescence probe–based DNase Activity Detection Kit and RNase Activity Detection Kit. These kits are designed to detect DNase and RNase contamination in laboratory environments, consumables, and raw materials, and for accurate quantification of residual nuclease activity in biopharmaceutical products, supporting effective process control and quality assurance.

The fluorescence probe–based DNase and RNase activity assays utilize nucleic acid substrates labeled with fluorophore–quencher pairs. In the absence of DNase or RNase activity, the substrate remains intact, and the fluorophore is maintained in close proximity to the quencher. Due to fluorescence resonance energy transfer (FRET), fluorescence emission is effectively quenched, resulting in minimal background signal.When DNase or RNase activity is present, the nucleic acid substrate is cleaved, causing separation of the fluorophore and quencher. This disruption of the FRET effect leads to a progressively increasing fluorescence signal. The rate of fluorescence increase is directly proportional to the enzyme concentration and catalytic activity in the sample.

Fluorescence intensity is measured using a microplate reader at excitation/emission wavelengths of 490/520 nm for RNase and 535/565 nm for DNase. Based on the fluorescence signal, the presence of DNase or RNase contamination in the sample can be accurately determined.