Protein

— High Quality Drug Target Proteins

Cell-free protein expression system

Cell-Free Protein Synthesis (CFPS) is an in-vitro recombinant protein expression technology. It is an in-vitro translation system that synthesizes target proteins in a way similar to a "one - pot" chemical reaction. This system directly utilizes cell lysates containing the natural cellular transcription and translation machinery. By adding the target gene and appropriate co-factors such as amino acids, energy substrates, and salts, the synthesis of the target protein can be achieved in vitro within a few hours. This system bypasses the complex processes of cell culture and expression optimization, improving the efficiency and flexibility of protein synthesis. It is suitable for the preparation of various types of proteins, especially for the expression and purification of hard-to-express proteins, toxic proteins, and difficult proteins.

Advantages

Simple and quick

Complete expression within 4 hours, and finish SDS-PAGE/FSEC, TSA, and bioactivity assays within 1 day. It's feasible to conclude all the screening and detection procedures, covering SDS - PAGE, FCT, FSEC, and enzyme activity assays.

High-throughput

It is applicable to a variety of expression vectors and host systems (E.coli, insects, mammalian cells). The templates are flexible, with PCR products, mRNA, and plasmids all being viable as expression templates. Moreover, expression screening can be carried out on multi-well plates, making it suitable for high-throughput proteomics research.

Wide applicability

The open-reaction system makes it easy to modify the reaction conditions. It is suitable for the expression of various proteins, including membrane proteins and toxic proteins, as well as multiple protein modifications, such as isotope labeling and non-natural amino acid labeling.
Biortus E.coli CFPS

Applications

  • 1
    High-throughput screening and protein preparation
  • 2
    Rapid preparation of in vitro recombinant proteins at the microgram to milligram level
  • 3
    Incorporation of NMR (15N and 13C) isotope labeling and non-natural amino acids
  • 4
    Rapid sample preparation for X-ray & Cryo-EM

Details of CFPS Service At Biortus

Steps Service Item Service Description Biortus Features Duration
01 Plasmid Construction Codon optimization and full-gene synthesis To improve protein expression level and solubility, we provide screening of various fusion tags. 1-2 Weeks
Construction of different fusion tags
02 Small-scale Expression and Optimization Screening and optimization of various vectors and different reaction conditions, detected by SDS PAGE and FSEC to determine the optimal reaction conditions Our cell-free expression system can perform high-throughput screening to provide the optimal protein expression conditions 1-3 Days
03 Scale-up Purification Expression in a scale-up system above the milliliter level is available. For membrane proteins, we can provide screening services for different detergents, liposomes, and nanodiscs. 3-5 Days
The target protein can be purified by various chromatographic methods such as AC, IEC and SEC etc.
04 Quality Control Check purity, concentration, etc., and provide various QC reports including LC-MS - 1 Day
20 Days

Experimental Procedure

  • DNA template

    Plasmids with T7 promoter

    -

    PCR products

    -

    mRNA

  • Protein expression

    “One Pot” reaction

    -

    High-Throughput Screening

  • Expression detection

    SDS-PAGE

    -

    WB/Protein Simple

    -

    FSEC

Examples

Expression of the green fluorescent protein sfGFP

Biortus E.coli CFPS can efficiently express various proteins. For example, when the components of the kit are mixed and reacted with the plasmid expressing green fluorescent protein (GFP) , the fluorescence emitted by GFP can be observed without ultraviolet light irradiation.

Labeling of Selenocysteine and Selenomethionine

Biortus E. coli cell-free protein expression system enables the seleno-Cys and seleno-Met proteins: Simply mix the target selenoamino acid with the kit components and the labeled protein can be obtained after several hours of incubation.

Protein

Protein expression