On-demand, cell-free biomanufacturing of conjugate vaccines at the point-of-care

Conjugate vaccines are among the most effective methods for preventing bacterial infections, representing a promising strategy to combat drug-resistant pathogens. However, existing manufacturing approaches limit access to conjugate vaccines due to centralized production and cold chain distribution requirements. To address these limitations, we developed a modular technology for<underline>i</underline>n vitrobioconjugate<underline>va</underline>ccine e<underline>x</underline>pression (iVAX) in portable, freeze-dried lysates from detoxified, nonpathogenicEscherichia coli. Upon rehydration, iVAX reactions synthesize clinically relevant doses of bioconjugate vaccines against diverse bacterial pathogens in one hour. We show that iVAX synthesized vaccines against the highly virulent pathogenFranciscella tularensissubsp.tularensis(type A) strain Schu S4 elicited pathogen-specific antibodies in mice at significantly higher levels compared to vaccines produced using engineered bacteria. The iVAX platform promises to accelerate development of new bioconjugate vaccines with increased access through refrigeration-independent distribution and point-of-care production.