Method for cutting polypeptides based on solid phase synthesis

Abstract

The invention relates to the field of biochemistry and molecular biology and provides a method for cutting polypeptides based on solid phase synthesis. The method comprises the following steps of: (1) preparing three cutting fluids with different concentrations; (2) selection of the cutting fluids: selecting different cutting fluids according to different amino acids in a polypeptide sequence; (3) controlling time and temperature of the cutting reaction, wherein the cutting time is between 2 hours and 2 hours and 45 minutes, and the cutting temperature is less than or equal to 25 DEG C; (4) carrying out the cutting steps: 1, pouring the polypeptide resin needed to be cut into a centrifuge tube and selecting the cutting fluid according to the different amino acids in the peptide sequence; 2, performing the cutting reaction according to the requirements of step (3); 3, separating the resin and the polypeptide from a sand-core funnel after the cutting reaction; 4, pouring into cold ether to precipitate the polypeptide, centrifuging, precipitating and washing with ether, and 5, removing ether to obtain the final product. The cutting method provided by the invention is environmental-friendly, can reduce damages to operators, reduce the cost and improve the efficiency of polypeptide cutting.

Description

technical field [0001] The present invention relates to the fields of biochemistry and molecular biology, specifically a cutting method based on solid-phase synthesis of polypeptides, which is a cutting method in which different cutting liquids are selected according to the amino acids in the peptide sequence and the polypeptide can be eluted method. Background technique [0002] Polypeptide is a compound formed by linking α-amino acids with peptide chains, and it is also an intermediate product of protein hydrolysis, referred to as peptide for short. The compound formed by the dehydration condensation of two amino acid molecules is called a dipeptide, and by analogy there are tripeptides, tetrapeptides, pentapeptides... all the way to nonapeptides. Usually, the compound formed by dehydration condensation of 10-100 amino acid molecules is called polypeptide. Their molecular weight is lower than 10,000Da (Dalton Dalton), can pass through a semi-permeable membrane, and will ...

AI Technical Summary

Problems solved by technology

[0017] (1) Hydrofluoric acid (HF) is a highly acidic and highly toxic substance. Once inhaled, HF gas will endanger the human body and even cause death. The overflow of HF gas will cause serious pollution to the environment.

[0018] (2) HF is highly corrosive, and the cutting of polypeptides cannot be carried out in glass instruments, and experiments must be carried out in instruments of special materials

[0019] (3) The operation of cutting polypeptides with HF is difficult, so it must be carried out in a fume hood capable of strong corrosion; the operator must wear protective equipment such as eye masks, gas masks, and corrosion-resistant rubber gloves

[0020] (4) The use of nitrogen to evaporate HF and DMS after cutting with HF increases the complexity of the operation

[0021] (5) There may be side reactions such as aminoacylation involved in the reaction