Abstract
The market of biomolecules with therapeutic scopes, including peptides, is continuously expanding. The interest towards this class of pharmaceuticals is stimulated by the broad range of bioactivities that peptides can trigger in the human body. The main production methods to obtain peptides are enzymatic hydrolysis, microbial fermentation, recombinant approach and, especially, chemical synthesis. None of these methods, however, produce exclusively the target product. Other species represent impurities that, for safety and pharmaceutical quality reasons, must be removed. The remarkable production volumes of peptide mixtures have generated a strong interest towards the purification procedures, particularly due to their relevant impact on the manufacturing costs. The purification method of choice is mainly preparative liquid chromatography, because of its flexibility, which allows one to choose case-by-case the experimental conditions that most suitably fit that particular purification problem. Different modes of chromatography that can cover almost every separation case are reviewed in this article. Additionally, an outlook to a very recent continuous chromatographic process (namely Multicolumn Countercurrent Solvent Gradient Purification, MCSGP) and future perspectives regarding purification strategies will be considered at the end of this review.
Highlights
Peptides are organic polymers composed of 2–50 amino acids linked to each other by means of covalent amide (=peptide) bonds
Peptides can be classified as endogenous, if they are synthesized inside the human body, or exogenous if they are introduced into the organism from an external source
Solid Phase Peptide Synthesis (SPPS), firstly developed by Merrifield [46], one N-protected amino acid reacts with the peptide chain, which is anchored to a solid support and, after that, the terminal amino acid is deprotected
Summary
Peptides are organic polymers composed of 2–50 amino acids linked to each other by means of covalent amide (=peptide) bonds. The literature about different kinds of bioactivity that peptides can exert on the human body is broad They can interact with different organs and systems, such as digestive, cardiovascular, nervous and immune systems. Peptides can be classified as endogenous, if they are synthesized inside the human body, or exogenous if they are introduced into the organism from an external source It is well-known that proteins acquired through food are an essential source of amino acids and it has been demonstrated that some specific portions of dietary proteins exert biological functionalities. The first type of impurities can be separated from the peptide of interest, e.g., through affinity chromatography, during the so-called capture step [25] In this chromatographic technique, the ligand binds to the target molecule, whereas the impurities are flushed through the column. Other processes involve to synthesize the peptide-chain starting from single amino acids, adding one amino acid at a time [18]
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