Method for cutting polypeptides based on solid phase synthesis

Abstract

The present invention relates to the fields of biochemistry and molecular biology. It is a polypeptide cutting method based on solid-phase synthesis. The steps include: (1) preparing three kinds of cutting fluids with different concentrations; (3) Cutting reaction time and temperature control, the cutting reaction time is 2 hours to 2 hours and 45 minutes; the cutting temperature is controlled at ≤25°C; (4) Cutting steps: ①Pour the polypeptide resin to be cut into a centrifuge tube, according to The amino acid in the peptide sequence is different and the cutting solution is selected; ②The cutting reaction is carried out according to the requirements of step ⑶; ③After the cutting reaction, the resin and the polypeptide are separated from the sand core funnel; Cleaning; ⑤ removing ether to obtain the cleaved polypeptide. The cleavage method of the invention is environmentally friendly, can reduce harm to operators, reduces costs, and improves the efficiency of polypeptide cleavage.

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