Commonly Used Protein Expression Systems

Pros & Cons & Applications

Creative BioMart

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Protein expression, a kind of molecular biology technology referring to the use of
model organisms such as bacteria, yeast, animal cells, or plant cells to express
foreign gene protein, plays an important role in the field of genetic engineering.

This post is to make a comparison about the advantages, disadvantages and

applications of some commonly used protein expression systems, including
Escherichia coli expression system, Pichia pastoris expression system,
Saccharomyces cerevisiae expression system, Mammalian expression system,
Baculovirus Cell expression System, stable cell line expression system, etc.

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1. Escherichia coli expression system

Advantages
High-level expression; Low cost; Simple culture conditions; Fast expression;
Extensible; Easy to manipulate; Easy to form disulfide bonds.

Disadvantages
Easy to form inclusion bodies; Ineffective in vitro folding; Different codon systems;
Few post-translational modifications; Endotoxin

Applications
Production of purified protein (structure, enzyme, drug discovery); Drug protein
production.

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2. Pichia pastoris expression system

Advantages
High-level expression; Low cost; Simple culture conditions; Fast expression;
Extensible; Suite for secretory protein or intracellular expression; Protein
secretion is efficient and allows simple purification; Extensive post-
translational modification; N-terminal glycosylation is superior to
Saccharomyces cerevisiae; No endotoxin.

Disadvantages
Use of methanol as an inducer has a certain hazard; Glycosylation is still
different from mammalian cells.

Applications
Production of purified protein (structure, enzyme, drug discovery)

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3. Saccharomyces cerevisiae expression system

Advantages
High-level expression; Low cost; Suitable for secretory protein or
intracellular; Simple culture conditions; Extensible; Effective protein folding;
No endotoxin.

Disadvantages
Secretory capacity may be lower than Pichia pastoris; Glycosylation is
different from mammalian cells; Glycosylation N-terminal glycosyl chain
structure has sensitization.

Applications
Production of purified protein (structure, enzyme, drug discovery)

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4. Mammalian cells expression system

Advantages
Higher-level expression; Moderate scalability; Suspension culture
characteristics of the cells can be produced on a large scale; Effective protein
folding; Suitable for secreting proteins; Full post-translational modification;
No endotoxin.

Disadvantages
Expensive medium; Complex growth conditions

Applications
Production of purified protein (structure, enzyme, drug discovery); Drug
protein production; Cellbased studies.

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 5. Stable cell line expression system

Advantages
Protein production lines can be obtained

Disadvantages
Production is slow (takes several months), especially in the case of selective
cloning, the ability to express in the passage of the process may decline odification;
No endotoxin.

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6. Baculovirus-Insect Cell expression Systems

Advantages
High-level expression (especially intracellular protein); Faster growth
rate; Effective cell folding; Moderate scalability; Extensive post-
translational modification; Glycosylation is similar to mammalian cells;
Relatively easy enzymatic de-glycosylation (beneficial for protein
structure assays); No endotoxin.

Disadvantages
Expensive medium; Need a lot of virus; Secretion pathway of the pro-
peptide is inefficient; Glycosylation is still different from mammalian
cells; Virus infection can lead to cell lysis and potential expression of
protein degradation.

Applications
Production of purified protein (structure, enzyme, drug discovery)

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