What Are the Methods for Deproteinizing Polysaccharides? Is Drying Necessary After Deproteinization?
Extracting polysaccharides from protein-rich biological samples presents a significant challenge in biological and biochemical research. Two commonly used methods for polysaccharide deproteinization are as follows:
Trichloroacetic Acid (TCA)/Acetone Precipitation Method
Proteins can be precipitated using TCA or a cold acetic acid-acetone mixture, allowing polysaccharides to remain in the supernatant. The precipitated proteins are then washed with cold acetone to remove residual polysaccharides.
Chloroform:n-Butanol (4:1) Extraction Method
In this widely used deproteinization method, the sample is extracted with a chloroform:n-butanol (4:1) mixture. Proteins dissolve in the organic phase, whereas polysaccharides remain in the aqueous phase.
These methods may require optimization depending on the specific properties of the sample and the target polysaccharide.
The necessity of drying polysaccharides after deproteinization depends on the intended downstream analysis or application. For instance, mass spectrometry analysis often requires drying, while in other cases, drying can facilitate long-term storage. However, it is important to note that some polysaccharides may undergo physical changes when completely dried.
To further purify polysaccharides, dialysis or filtration can be employed to remove low-molecular-weight impurities, followed by lyophilization to obtain a dry powder. This process yields polysaccharides that are easier to store and transport. If a liquid form is required for subsequent experiments, the dried polysaccharides can be rehydrated in an appropriate solvent, such as purified water or a buffer solution.
MtoZ Biolabs, an integrated chromatography and mass spectrometry (MS) services provider.
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