Syngenta: Multiplex protein screening enables success with hard-to-express agricultural proteins

Syngenta trialed eProtein Discovery™ to screen diverse soluble and membrane agricultural proteins in parallel. This case study is based on a talk given by Syngenta at the 17th Annual PEGS Europe, Protein & Antibody Engineering Summit earlier this year.

 

Why crop protection advancements require proteins

To meet the demands of crops for today’s and future populations, agricultural companies are working to reduce the environmental impact of food production by increasing yield on existing land, improving tolerance to stressors such as drought or heat, developing safer tools to protect crops, and boosting nutritional value. 

 

Syngenta, a global leader in crop protection and seeds, is working towards safeguarding agricultural crops by developing chemical and biological protection technologies and improved seed varieties to increase yield, reduce environmental impact, and manage resistance. At the heart of this work are proteins.

 

Protein-based studies are essential for understanding a molecule’s mode of action, mapping structure–activity relationships, anticipating how microbes and pests might evolve resistance, and assessing possible off-target effects on human proteins (Figure 1). That means scientists need access to many different proteins from plants, microbes, and insects (Figure 2). Studying complex crop systems also means that scientists need to express difficult proteins such as membrane proteins or toxins. 

 

 

For Syngenta, this diversity makes protein production particularly challenging, especially in cell-based systems optimized for a single host.

 

“We’re always looking for tools and ideas to diversify our protein expression toolkit,” said Shradha Singh, Protein Science Lead, Bioscience at Syngenta. “Cell-free has been on our radar for quite some time.”

 

 

 

 

 

 

 

Breaking away from typical protein production workflows

 

One reason for turning to cell-free protein expression is that they can overcome the challenges of typical protein production workflow where scientists design genes, express them in cells, purify the proteins, and perform quality checks before moving into  downstream assays. These workflows can suffer from low expression levels, protein instability and insolubility, and trouble scaling up to quantities needed for structural biology or functional assays. Re-optimizing constructs and conditions can add weeks or months to a project, before you obtain the critical protein that enables development of crop protection technologies. 

 

Given Syngenta’s need to express numerous types of proteins, we were excited to partner with them on a demo of our eProtein Discovery™ system, an automated digital microfluidics system that screens and expresses soluble and membrane proteins from a number of biological sources using cell-free reagents. The goal of eProtein Discovery is to make screening protein variants and expression conditions faster with the ability to produce assay-ready proteins in 48 hours. The Syngenta team presented their findings at the recent 17th Annual PEGS Europe, Protein & Antibody Engineering Summit earlier this year.

 

Case study: Multiplex protein screening for diverse agricultural proteins

 

In this demo, Syngenta expressed 9 different soluble proteins of plant, fungal, and bacterial origin, and 9 membrane proteins of insect, fungal, and plant origin under various conditions and with different tags within 24 hours. The team then compared yields from cell-free expression using eProtein Discovery with yields from their in-house cell-based expression workflow.

 

Soluble proteins

For soluble proteins, Syngenta found that, for the most part, yields from cell-free expression matched that from cell-based expression. One bacterial pore forming protein showed higher yield from cell-free expression compared to cell-based. After scaling up the top 3 soluble proteins, the team saw that the expression results remained comparable with cell-based expression. Xinghao Zhou, Senior Protein Scientist at Syngenta, referred to this as “a very exciting result.” Historically, cell-free systems have often resulted in lower yields compared to cell-based expression.1

 

Membrane proteins

For membrane proteins, the team tested expression of insect transporters, an ion channel, an enzyme, a receptor, and a plant toxin protein, and took the top four proteins for scale-up. After scale-up, these proteins reached yields of 0.37 mg/mL to 0.6 mg/mL, similar to what the team observed for their in-house cell-based expression, but in a much faster time. The screen quickly identifies which proteins are promising within 24 hours, offering assay-ready protein in 48 hours.

 

 

Crucially, the plant toxin protein, which the team had not been able to express successfully in a variety of host expression systems, had a yield of 0.4 mg/mL using eProtein Discovery. Additionally, a quality control of the insect transporters generated revealed successful binding through thermal shift assays following ligand-binding.

 

This demo produced what Singh called “encouraging results for Syngenta’s diverse and challenging targets.” It demonstrated the versatility of eProtein Discovery by successfully expressing a variety of proteins from different biological sources, including a protein that couldn’t be expressed in cell-based systems, and by achieving higher yields with novel tags in insect toxins. The Syngenta team is currently working on optimizing lipid compositions during membrane protein expression for activity.

 

“eProtein Discovery is really a hands-off, streamlined screening system that you can walk away from and come back in 48 hours to look at your protein results,”

said Singh.

 

 Start your protein screening here

 

References

1. Hunt, A. C., Rasor, B. J., Seki, K., Ekas, H. M., Warfel, K. F., Karim, A. S., & Jewett, M. C. (2025). Cell-Free Gene Expression: Methods and Applications. Chemical reviews, 125(1), 91–149. https://doi.org/10.1021/acs.chemrev.4c00116