APPLICATIONS OF TECHNOLOGY:
- Biomanufacturing of enzymes
- Enzyme research
ADVANTAGES:
- Rapid discovery of the function of genes
- Capable of multiplexing
- Ultra-high-throughput
ABSTRACT:
Researchers at Berkeley Lab led by Trent Northen have developed the ARGO (Assignment of the Reactions of Gene products in Organisms) platform, an efficient and scalable technology providing insights into gene function by quickly and reliably determining the function of putative proteins to understand microbial metabolism.
ARGO is based on using mass spectrometry to read out the identity, quantity and activity of gene products. Specifically, a peptide barcode that can be detected along with reaction substrates and products, using mass spectrometry, encodes the identity of the genes and is used to quantify the genes. Genes are linked via the sequence for a protease labile site such that after expression, protease treatment produces the enzymes and two peptides. One peptide is a barcode that encodes the identity of the expressed enzyme, the second peptide is a conserved quantification peptide. This mixture then can be screened for activities against a range of substrates, co-factors and reaction conditions to discover the activities of the enzyme. The resulting reaction products and peptide barcodes are identified using mass spectrometry. Using an internal standard, the quantity of expressed protein can be determined to estimate the specific activities of the enzyme.
The ARGO platform enables combinatorial experimental designs, allowing for multiplexing and ultra high-throughput. Where spectrometry-based approaches are hindered by low-throughput and a limited ability to multiplex, the ARGO platform enables rapid discovery of the function of genes.
DEVELOPMENT STAGE: Proven principle
STATUS: Published U. S. Patent Application 15/663,528 (Publication No. 2018/0030531). Available for licensing or collaborative research.
SEE THESE OTHER BERKELEY LAB TECHNOLOGIES IN THIS FIELD:
Long-range Barcode Labeled Sequencing for Efficient Genome and Metagenome Assembly IB-3019