Polypeptide solid phase synthesis history.

In 1963, R.B. Merrifield created a solid-phase synthesis method that fixed the C end of the
amino acid
to an insoluble resin, and then in turn indentated the amino acids on the resin, prolonging the solid-phase synthesis of peptide chains and synthesizing
proteins
, in which the solid phase method can be purified by simply washing the resin after each step of reaction. Overcoming the difficulty of purification of every step of the product in the classical liquid phase synthesis method, it lays the foundation for automatic synthesis of peptides. To this end, Merrifield won the 1984 Nobel Prize in Chemistry.
today, the solid phase method has been greatly developed. In addition to the Boc method established by Merrifield (Boc: Schotinoxyl), the Fmoc solid phase method (Fmoc:9- methoxycoxyl) has been developed. Various peptide autosynthetic meters based on these two methods have also appeared and developed one after another, and are still being modified and perfected.
's Boc synthesis method, established by Merrifield, is a α-amino protection base using TFA (tefluoroacetic acid) removable boc, and side chain protection using penicillin. Synthesis of a Boc-amino acid derivative co-priced to the resin, with TFA to remove Boc, with triethylamine and free amino ends, and then through Dcc to live, couple the next amino acid, and finally depulption using HF or TFMSA (TFMSA) method. The Boc method has been successfully synthesized into many biological molecules, such as active enzymes,
growth factors
artificial proteins, etc.
peptides
biologically active substances involving various cellular functions in the organism. It is a class of
compounds
with molecular structure between amino acids and proteins, which are combined by peptide bonds in a certain order of amino acids. Up to now, more than 100 kinds of peptides present in the human body have been discovered and isolated, and the research and utilization of peptides has seen an unprecedented prosperity.
synthesis of peptides is not only of great theoretical significance, but also of great application value. The structure of a new peptide can be verified by the total synthesis of peptides, new peptides can be designed to study the relationship between structure and function, important information can be provided for the mechanism of polypeptide biosynthetic reaction, model enzymes are established and new peptide drugs are synthesized.
chemical synthesis techniques of polypeptides have matured both liquid and solid phase methods. In recent decades, solid-phase synthesis polypeptides have become a common method of peptide synthesis with their outstanding advantages of time-saving, labor-saving, material-saving, computer-friendly and popularization, and extended to other
organized
areas such as
nucleotides
synthesis. This paper summarizes the basic principles and experimental process of solid phase synthesis, analyzes its current situation and looks forward to the future development trend.
Since Merrifield developed the successful method of solid-phase polypeptide synthesis in 1963, after continuous improvement and improvement, solid-phase method has become a common technique in polypeptide and protein synthesis today, showing the advantages of classical liquid-phase synthesis.
The basic principle is: first the hydroxyl of the hydroxy-end amino acids to be synthesized peptide chain is connected with an insoluble polymer resin structure with an insoluble polymer resin, and then the amino acids combined on the solid-phase carrier are used as amino groups to be stripped of amino protection and react with excessive active pyrethroides, and the peptide chain is attached.
Repetition (shrinking→ washing→ to protect→ and washing →the next round of shrinking) operation, to achieve the length of the peptide chain to be synthesized, and finally the peptide chain from the resin cracked down, after purification and other treatment, that is, the desired peptide. Among them α-amino is protected by BOC (Shoddyxycoxyl) called BOC solid-phase synthesis, and α-amino is protected by FMOC (9- methoxycoxyl) called FMOC solid-phase synthesis.
Principle of solid-phase synthesis
Polypeptide synthesis is a process of repeated addition of amino acids, and solid-phase synthesis is generally synthesized from the C end (the base end) to the N end (the amino end). In the past, peptide synthesis was done in a solution called liquid phase synthesis. Now more solid-phase synthesis method, which greatly reduces the difficulty of purification of each step of the product. In order to prevent the occurrence of side reactions, the side chains of the amino acids participating in the reaction are protected. The base end is free and must be active before reaction. There are two methods of chemical synthesis, namely Fmoc and tBoc. Since Fmoc has many advantages over tBoc, it is now mostly composed of Fmoc, as shown in Figure:
specific synthesis consists of the following cycles: 1, to protect: Fmoc-protected columns and monobodies must be removed with an alkaline solvent (piperidine) to remove amino protection groups., activation, and crosslinking: the carboxyl of the next amino acid is activated by an activator. The active monosome is interlinked with the free amino reaction to form a peptide bond. In this step, a large number of
reagents are used
drive the reaction to complete. Loop: These two-step reactions are repeated until the composition is complete., equipe and de-protect: peptides are removed from the column and their protective groups are de-protected by a de-protective agent (TFA).
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