Biological Additive Manufacturing Capabilities for Enabling Rapid Response to Emerging Pathogens

Project Overview

The ability to perform high throughput screening is critical for developing therapeutics for new and reemerging threats and pathogens. Strategies for increasing the efficiency and speed of this screening are valuable for testing candidate therapeutics combinatorial/random compounds and from computational efforts. In this work we advanced strategies for combining steps in the screening process. In general, the production of proteins is a separate step from a screening step based on measuring interactions between proteins. This is especially true with in vitro techniques. As part of this LDRD we were able to demonstrate that cell-free process can be used to produce dozens of recombinant proteins of interest for studying affinity interactions. This automated process is compatible with microfluidic chips for generating capsules that are compatible for delineating pM affinities of antibody derived interactions simultaneously using fluorescent spectroscopy. This includes designing a device for potentially automating the entire process to potentially process thousands to millions of protein or small molecule samples for production as well as analysis per week.

Mission Impact

This project builds on Lawrence Livermore National Laboratory's (LLNL) Core Competencies in Bioscience and Bioengineering, as well as High Performance Computing. It responds to the Laboratory's mission in Threat Preparedness and Response by building a new avenue of research in synthetic biology combined with advanced single molecule detection and microfluidics to develop a novel benchtop device for protein production (104 -105) and analysis of >106 assays. These integrated technologies fill a gap in quantitative predictive biology by providing instrumentation and protein reagents to predict and develop pathogen countermeasures, potentially in under a week. This work will also support future mission space under the JPEO GUIDE and Bioresilience programs.

Publications, Presentations, and Patents

Chao Liu, Steven Hoang-Phou, Congwang Ye, Matthew Laurence, Bonnee Rubinfeld, Byron Dillon Vannest, Thomas Desautels, William Smith, Erika Fong, Nicholas Watkins, Sean Gilmore, Maxim Shusteff, Ted Laurence, Matthew Coleman, "Fluorescence correlation spectroscopy of proteins expressed inside microcapsules Developing a high throughput cell-free system for generating and characterizing protein-based countermeasures" (Presentation, Biophysical Society, San Francisco, CA, Feb 2022).

Congwang Ye, Steven Hoang-Phou, Nikki Hammond, Chao Liu, Erika Fong, Nicholas Watkins, Matthew Laurence, William Smith, Sean Gilmore, Ted Laurence, Daniel Faissol, Matthew Coleman, "One-step microencapsulation of cell-free lysate for improved protein synthesis and characterization" (Presentation, Protein Society, San Francisco, CA, July 2022).

Matthew Coleman, "Nanoparticle-technology to support functional membrane bound receptors" (Invited Oral Presentation, Saint Jude Children's Research Hospital, Memphis, TN, 2021).

Matthew Coleman,"Biologically inspired nanoparticles as an engineered platform for biotechnology" (Invited Oral Presentation, Biotechnology Seminar Series, Georgetown University, Washington, D.C. 2021). 

Matthew Coleman,  "Development and applications of cell-free protein expression for building high-throughput pipeline for functional characterization of proteins" (Invited Oral Presentation, HHS BARDA DRIVe Tuesday Talk Series, Virtual, 2023).

Matthew Coleman,"Design of Natural and Synthetic Nano-Scaffolds for Antigen Biochemistry and Antigenicity Testing" (Invited Oral Presentation, KEMP Proteins, Washington, D.C., 2023). 

Matthew Coleman, "Applying nanodisc technologies for de novo cell-free synthesis of membrane proteins" (Invited Oral Presentation, Protein Engineering Summit (PEGS) Difficult to Express Proteins Conference, Boston, MA, 2023).  

Patent Application IL-13845P1 - Automated High-Throughput Screening Platform, LLNL.

Laurence, M.J., Carpenter, T.S., Laurence, T.A., Coleman, M.A., Shelby, M. and Liu, C. (2022). "Biophysical characterization of membrane proteins embedded in nanodiscs using fluorescence correlation spectroscopy." Membranes. 12(4), 392; doi.org/10.3390 LLNL-JRNL-836103