Journal: Methods and Protocols

Article Title: A User’s Guide to Cell-Free Protein Synthesis

doi: 10.3390/mps2010024

Figure Lengend Snippet: Comparison of protein yields across cell-free platforms. The volumetric yield of each platform is reported for batch reactions producing GFP. Platforms that report volumetric yield for reporter proteins luciferase (*) or ChiAΔ4 (**) are indicated. Information for batch mode protein yields of the Arabidopsis and Neurospora crassa platforms was not found. Yields were obtained from the following sources: E. coli , wheat germ , Vibrio natriegens , Leishmania tarentolae , tobacco , HeLa , Pseudomonas putida , Streptomyces , Bacillus megaterium , Chinese hamster ovary , insect , Bacillus subtilis , yeast , archaeal , and rabbit reticulocyte .

Article Snippet: Currently commercial kits exist for E. coli (New England Biolabs, Promega, Bioneer, Qiagen, Arbor Biosciences, ThermoFisher, Creative Biolabs), rabbit reticulocyte (Promega, Creative Biolabs), wheat germ (Promega, Creative Biolabs), Leishmania tarentolae (Jena Bioscience) , insect (Qiagen, Creative Biolabs), Chinese hamster ovary (Creative Biolabs), HeLa (ThermoFisher, Creative Biolabs), and plant cells (LenioBio).

Techniques: Comparison, Luciferase

Journal: Methods and Protocols

Article Title: A User’s Guide to Cell-Free Protein Synthesis

doi: 10.3390/mps2010024

Figure Lengend Snippet: Comparison of growth methods for high adoption platforms. We have assembled the major growth methodologies for each of the high adoption platforms to give users an idea of the relative differences between them. These are not the only techniques that have been used for growth for each platform, but they are generally representative of the methods.

Article Snippet: Currently commercial kits exist for E. coli (New England Biolabs, Promega, Bioneer, Qiagen, Arbor Biosciences, ThermoFisher, Creative Biolabs), rabbit reticulocyte (Promega, Creative Biolabs), wheat germ (Promega, Creative Biolabs), Leishmania tarentolae (Jena Bioscience) , insect (Qiagen, Creative Biolabs), Chinese hamster ovary (Creative Biolabs), HeLa (ThermoFisher, Creative Biolabs), and plant cells (LenioBio).

Techniques: Comparison, Solvent, Hood, Cell Culture

Journal: Methods and Protocols

Article Title: A User’s Guide to Cell-Free Protein Synthesis

doi: 10.3390/mps2010024

Figure Lengend Snippet: Comparison of extract preparation methods for high adoption platforms. We have assembled the major extract preparation methodology for each of the high adoption platforms to give users an idea of the relative differences between them. These are not the only techniques that have been used for extract preparation for each platform, but they are generally representative of the methods.

Article Snippet: Currently commercial kits exist for E. coli (New England Biolabs, Promega, Bioneer, Qiagen, Arbor Biosciences, ThermoFisher, Creative Biolabs), rabbit reticulocyte (Promega, Creative Biolabs), wheat germ (Promega, Creative Biolabs), Leishmania tarentolae (Jena Bioscience) , insect (Qiagen, Creative Biolabs), Chinese hamster ovary (Creative Biolabs), HeLa (ThermoFisher, Creative Biolabs), and plant cells (LenioBio).

Techniques: Comparison, Lysis, Concentration Assay, Sterility, Centrifugation, Saline, Disruption, Filtration, Extraction

Journal: Methods and Protocols

Article Title: A User’s Guide to Cell-Free Protein Synthesis

doi: 10.3390/mps2010024

Figure Lengend Snippet: Comparison of cell-free protein synthesis reaction setup for high adoption platforms. This table is intended to help users understand major differences between setups for various high adoption platforms, namely whether reactions are generally coupled or uncoupled, what energy systems are typical, and what temperatures the reactions are run at. These are not the only setups that have been used for successful cell-free protein expression in each platform, but they are generally representative of the reagents, concentrations, and temperatures used for each platform.

Article Snippet: Currently commercial kits exist for E. coli (New England Biolabs, Promega, Bioneer, Qiagen, Arbor Biosciences, ThermoFisher, Creative Biolabs), rabbit reticulocyte (Promega, Creative Biolabs), wheat germ (Promega, Creative Biolabs), Leishmania tarentolae (Jena Bioscience) , insect (Qiagen, Creative Biolabs), Chinese hamster ovary (Creative Biolabs), HeLa (ThermoFisher, Creative Biolabs), and plant cells (LenioBio).

Techniques: Comparison, Expressing, Maltodextrin, In Vitro, Purification

Journal: Heliyon

Article Title: Immunoproteomic approach identifies a putative virulence chaperone DnaK protein as a candidate diagnostic marker and therapeutic target for Pythium insidiosum infection

doi: 10.1016/j.heliyon.2025.e42487

Figure Lengend Snippet: Fig. 2. SDS-PAGE and Western blot analysis of P. insidiosum’s synthesized proteins. Nineteen coding sequences of the recruited target proteins S01- S21 (except S03 and S04) serve as a protein synthesis template in the CFPS reaction, and the resulting products are checked for their presence using SDS-PAGE stained with Coomassie blue dye (A) and their immunoreactivity using Western blot analysis probed against the mouse anti-6x-His tag antibody (B). Green fluorescent protein (GFP) is a positive control for the CFPS reaction. An arrowhead indicates a target synthesized protein. The molecular weight markers (M) are shown on the left of each figure. The original unprocessed images of Fig. 2 are available as supplementary files: Original_Fig2A_S01-02_S11-S14_GFP.tif, Original_Fig2A_S05-06.tif, Original_Fig2A_S07-10.tif, Original_Fig2A_S015-21.tif, Original_Fig2B_S01- S02_S11-S14_GFP.tif, Original_Fig2B_S05-06.tif, Original_Fig2B_S07-10.tif, and Original_Fig2B_S015-21.tif.

Article Snippet: Up to 16 different proteins can be synthesized simultaneously using a second-round PCR product as a template for protein synthesis and a commercial CFPS kit from Bioneer with an automated protein synthesis machine from the same company.

Techniques: SDS Page, Western Blot, Synthesized, Staining, Positive Control, Molecular Weight