In RP HPLC, one of the most common separation methods used to measure purity, strength, dosage, etc, a protein would be put into 0.1% (1000 ppm) TFA (Trifluoroacetic acid), what do you suppose this does to the protein in many cases? (Pick the BEST answer). somewhat denaturing to very denaturing oxidizes cysteine ionizes acid and base groups
Proteins
We generally tend to think of proteins only from a dietary lens, as a component of what we eat. However, they are among the most important and abundant organic macromolecules in the human body, with diverse structures and functions. Every cell contains thousands and thousands of proteins, each with specific functions. Some help in the formation of cellular membrane or walls, some help the cell to move, others act as messages or signals and flow seamlessly from one cell to another, carrying information.
Protein Expression
The method by which living organisms synthesize proteins and further modify and regulate them is called protein expression. Protein expression plays a significant role in several types of research and is highly utilized in molecular biology, biochemistry, and protein research laboratories.
High Performance Liquid Chromatography (HPLC) is biochemical separation method for organic molecules or solutes of a compound solution depending on the period of their respective interaction with the solid matrix. There are two separate phases in the matrix, one is mobile phase, which is liquid, another one is solid stationary phase. The mixture, whose compounds are to be separated are called analytes. Analyte compounds are carried by the fluid phase through the solid stationary matrix. The separation of the molecules depend on their affinity to the stationary matrix, for example if they are strongly bonded by covalent interaction to the solid phase, due to higher affinity, the velocity of the analyte's movement is slower than the analyte which do not bond or fix on the solid matrix, and flow with the mobile phase due to its higher affinity for the mobile phase. The velocity of the movement of the analyte compounds depend on their polarity, hence their interaction with the non-polar mobile phase. If the organic molecules are non-polar they move faster through the solid stationary phase. Proteins are organic molecules which are often separated by chromatographic technique as they have different side chains which are acidic and or basic in nature.
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