Bacterial reagents: stable, low-cost, readily available reagents with no purification requirements

Problem

Most molecular biology techniques commonly used in research, biotechnology, healthcare, and education rely heavily on purified functional protein reagents. However, purification of these protein reagents requires substantial investment of time, expertise, equipment, and infrastructure. As a result, this significantly limits the availability of these reagents to resources primarily with means of both accessibility and affordability. In addition to the investments in equipment, the costly prices of these purified reagents are often a roadblock for research conducted in laboratories in poor or remote settings. In order to enhance the availability of molecular biology reagents worldwide, there needs to be methodologies developed that simplify reagent production. This will make cellular reagents significantly cheaper to produce, store, distribute, and use for both educational and research purposes.

Solution

Researchers at The University of Texas at Austin have developed a method that simplifies reagent production by eliminating protein purification entirely. Instead, this new method uses lyophilized bacteria1. Now molecular biology techniques previously requiring costly and labor-intensive purified proteins, can simply be replaced by an optimal amount of corresponding rehydrated, lyophilized cellular reagents. To prove the feasibility of their approach, the researchers developed several lyophilized DNA polymerases and a thermostable reverse transcriptase. These novel, lyophilized reagents performed on par with their purified reagents counterparts in amplification efficiency, detection limits, and time across multiple analytical procedures. Additionally, the researchers demonstrated how their method is optimal for one-pot reactions through successful Gibson assemblies and most recently, incredible detection of SARS-Cov-2 virions in crude human saliva through loop-mediated isothermal amplification (LAMP) assays2. These lyophilized reagents are especially useful for biologists in resource-poor or educational settings, as they enable cost-efficient and fast results with with minimal training and resources required. This new method can revolutionize the supply chain and economics for molecular biology reagents.

References

1.  Bhadra S, Pothukuchy A, Shroff R, Cole AW, Byrom M, Ellefson JW, et al. (2018) Cellular reagents for diagnostics and synthetic biology. PLoS ONE 13(8): e0201681. https://doi.org/10.1371/journal.pone.0201681

2.  Sanchita Bhadra, Timothy E. Riedel, Simren Lakhotia, Nicholas D. Tran, Andrew D.Ellington. High-surety isothermal amplification and detection of SARS-CoV-2, including with crude enzymes. bioRxiv 2020.04.13.039941; doi: https://doi.org/10.1101/2020.04.13.039941