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שלמות mRNA

שלמות mRNA

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שלמות mRNA

The Significance of mRNA Integrity Analysis

mRNAs are recognized for their inherent instability and are susceptible to fragmentation in the presence of external factors, particularly during in vitro transcription (IVT) and purification processes, where ubiquitous nucleases pose a challenge. Evaluating mRNA integrity is crucial in the subsequent development of mRNA-based drugs, ensuring both safety and product availability.

Yaohai-BioPharma provides comprehensive mRNA integrity testing services, employing methods such as agarose gel electrophoresis (AGE), capillary electrophoresis (CEG), and high-performance liquid chromatography (HPLC), adhering to rigorous quality standards for both research and clinical trials.

דרישות רגולטוריות לשלמות mRNA

According to the WHO regulatory considerations, “the integrity of the structure of the mRNA is considered to be a critical quality attribute for the release of the mRNA. Thus, control is needed for mRNA integrity, percentage intact mRNA, percentage mRNA fragments and so on.”

שיטה אנליטית

אָנָלִיזָה

שיטות

mRNA integrity

AGE, Agarosegel electrophoresis

CGE,אלקטרופורזה נימית

CGE-LIF, Capillary gel electrophoresis with light induced fluorescence detection

mRNA Integrity by Agarose Gel Electrophoresis

Agarose gel electrophoresis is a widely employed technique for evaluating the integrity of total RNA. This involves running the RNA sample on a denaturing agarose gel, stained with either ethidium bromide or SYBR Green. The denaturing gel, composed of agarose, facilitates the separation of RNA based on size when an electric current is applied. As the negatively charged RNA migrates towards the positive end of the gel, smaller fragments move more rapidly, resulting in a size-based sorting of RNA molecules.

mRNA Integrity by Capillary Gel Electrophoresis

Capillary gel electrophoresis (CGE) stands as a high-resolution analytical approach pivotal for gauging the integrity of RNA molecules by length. This assessment holds significance for quality assurance, potency comprehension, and optimization of manufacturing processes. Similar to agarose gel electrophoresis, CGE utilizes an electrical current to segregate RNA fragments based on their charge and size. In CGE, a capillary is filled with a separation gel matrix infused with a fluorescent dye. The movement of RNA along the capillary is contingent on its size, and detection is achieved using ultraviolet (UV) or light-induced fluorescence (LIF) detectors.

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