Throughout downstream processing, various analytical methods must be used to evaluate the effectiveness of unit operations. Provide a method or methods (if more than one is required) for each scenario. Determination of the pH that the protein product carries a net neutral charge: Protein purity after each separation method: Characterize proteins of a complex lysate based on charge and molecular weight: Determine if a protein’s activity has been impacted by process operations:
Enzyme kinetics
In biochemistry, enzymes are proteins that act as biological catalysts. Catalysis is the addition of a catalyst to a chemical reaction to speed up the pace of the reaction. Catalysis can be categorized as either homogeneous or heterogeneous, depending on whether the catalysts are distributed in the same phase as that of the reactants. Enzymes are an essential part of the cell because, without them, many organic processes would slow down and thus will affect the processes that are important for cell survival and sustenance.
Regulation of Enzymes
A substance that acts as a catalyst to regulate the reaction rate in the living organism's metabolic pathways without itself getting altered is an enzyme. Most of the biological reactions and metabolic pathways in the living systems are carried out by enzymes. They are specific for their works and work in particular conditions. It maintains the best possible rate of reaction in the most stable state. The enzymes have distinct properties as they can proceed with the reaction in any direction, their particular binding sites, pH specificity, temperature specificity required in very few amounts.
Throughout downstream processing, various analytical methods must be used to evaluate the effectiveness of unit operations. Provide a method or methods (if more than one is required) for each scenario.
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Determination of the pH that the protein product carries a net neutral charge:
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Protein purity after each separation method:
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Characterize proteins of a complex lysate based on charge and molecular weight:
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Determine if a protein’s activity has been impacted by process operations:
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Assess total protein in rice flour (solid material).
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Visualize antibody fragments (heavy chain and light chain) and compare molecular weights.
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Quick determination of the concentration of a purified sample of monoclonal antibody.
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Determine the concentration of monoclonal antibody in clarified extract.
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Study aggregation of a protein product without disruption of tertiary protein structure.
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Characterize phenolics (secondary metabolites and impurities produced by green plant tissue) by identifying phenolic species and relative concentrations.
11. Detect protein variants that differ by a substitution of two amino acids.
12. Quantify the level of endotoxin in the final purified product.
13. Quantify bacterial contamination in your final formulated product.
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