How Much Do You Know about Cell-free Protein System?

Posted by Ellen Burns on February 1st, 2021

As a kind of in vitro synthetic protein expression method, the cell-free protein expression system has the advantages of high efficiency and speed, and its operation is very simple. In biological understanding, this method has a great effect and its application value is very wide. The cell-free protein expression system has become the focus of protein drug production. In the process of screening high-throughput drugs, it provides ideas and new methods for solving problems. At the same time, many people have achieved good results in the field of biopharmaceuticals, and related researchers have continued to work hard to promote the development and progress of the industry and show good development potential.

With the progress of society and the development of science and technology, more and more people have studied protein, and its research has developed to the cutting-edge field, which has received much attention. Cell-free protein expression systems have also received much attention. At present, the research of this system has gone into deeper fields, and the cost has become lower and lower. Compared with the traditional in vivo expression system, the cell-free protein system has the advantage of not being limited by cell physiology, and the cell-free protein system is based on cell extracts, which can express a large number of toxic proteins without inhibition by cellular proteases, and this time is very short, does not require complex treatment downstream, and plays an increasing role in the field of biopharmaceuticals. At the same time, complex proteins have also begun to be reflected in biopharmaceutical engineering, reflecting advantages such as toxic proteins and membrane proteins. However, due to the natural protein half-life, water solubility is very poor. In order to realize its expression, realize protein stability and exert its potential, it has become a hot topic in today's society.

Classification of expression system of acellular protein

At present, cell-free protein systems are mainly divided into two expression system types, which are prokaryotic and eukaryotic. The two have different characteristics and systems, and the needs in the research process are also different, so, it is necessary to select the appropriate needs.

Prokaryotic system

Since the study of prokaryotic expression systems, the realization of its expression process has utilized the extracts of biological cells, of which E. coli is very commonly used. It is very well tolerated. And the expression system established on this basis has great advantages. The expression of specific target proteins requires the expression of target proteins based on marker proteins and protease inhibitors. However, it has certain disadvantages, which is that it is very difficult to correctly fold after translation. However, E. coli has low cost, high expression efficiency, and high application value.

Eukaryotic system

The relationship between eukaryotic and prokaryotic systems is relative. Eukaryotic systems utilize extracts from eukaryotes during expression. Among them, rabbit reticulocytes, malt extracts are common. The eukaryotic expression system is not inhibited, so the expression is very efficient. In the process of eukaryotic expression system, in order to better realize gene recombination and more stable expression, it is necessary to linearize the circular template and ensure the enhancement of efficiency. Relevant studies have shown that E. coli and malt extract systems have high synthetic amounts and can be high-throughput studied.

Application in the field of biopharmaceuticals

Cell-free protein expression systems have long been costly to produce and inefficient, and these shortcomings have caused some limitations to their development. Its development value is very high in the field of biopharmaceuticals. With the continuous development and progress of biopharmaceutical technology, the relevant energy is also improving, and the preparation of cell extract process is also continuously developing, realizing the high-throughput expression of protein in vitro, and playing a role in promoting the development of the application field of cell-free protein system. In the field of biopharmaceuticals, cell-free protein systems play a very important role and solve many problems.

Introduction of unnatural amino acids

The introduction of unnatural amino acids into the protein sequence is a way to modify protein engineering, in order to give chemical modification of proteins and provide genes for proteins. In the traditional way, inhibitory tRNAs are added to recognize the termination code, and then nonsense mutated genes are carried as templates to introduce amino acids at specific sites. Relevant studies have found that unnatural amino acid technology is very effective, can achieve the specific modification of protein drugs, and give them certain pharmacological characteristics, such as can improve the kinetic stability of drugs. At present, if unnatural amino acid substrates are added to the cell-free protein system, an efficient reaction system can be established and the target protein can be selectively modified.

Large-scale production of recombinant protein drugs

The cell-free protein expression system can be derived from E. coli, and this expression system is continuously developed and continues to be developed and produced after the translation of complex structural membrane proteins. Under this system, the expression of luciferase has been realized and the modification of glycosylation has been realized, which is of great research value and significance. And related studies have also demonstrated that cell-free protein expression technology is very promising in recombinant proteins.

High-throughput expression of proteins

It is very important to elaborate the relationship between the realization process of protein gene coding and structure and function, and it has become one of the problems to be discussed in the development of today's society. In the context of this era, high-throughput protein expression technologies are very important. Cell-free protein expression systems are also very important and are favored by academic researchers at this stage. Compared with the traditional expression system, the cell-free protein system has certain advantages in expression, and it eliminates the molecular cloning process and can use PCR as a template. For some complex proteins, they cannot be expressed in vivo and there is no way to achieve protein folding. For these advantages to be fully exerted and utilized, researchers are very flexible and easy to study in the process of protein research. In addition, the cell-free protein system can efficiently prepare protein chips. It can further realize protein immobilization and has no way to purify soluble proteins, which saves money. At present, many studies have shown that protein chip technology has been widely used in clinical diagnosis and plays an effective role.

Cell-free protein expression system is a rapid and efficient method for protein synthesis in vitro. Compared with traditional in vivo expression, it can make up for its shortcomings and realize the expression in vitro. Therefore, it has great development potential in the field of biopharmaceuticals and has a wide range of application value. However, there are still many shortcomings in the development of cell-free protein system at present, and it is necessary to carry out in-depth research to deeply explore its reaction system, to deeply mine the synthesis of the system, and to better promote its development.