High throughput Formulation Automated Screening Platform for Nucleic Acid Encapsulated LNPs

Components of Nucleic Acid Encapsulated LNPs Genetic Materials Lipid Components Cationic/ionizable lipid Helper lipid PEGylated lipid SiRNA PreciGenome

Lipid Components and Functions Helper Lipids Cationic/Ionizable Lipids • Increase nucleic acid encapsulation rate Critical for endosomal escape Increase transfection efficiency DOPE –facilitate fusion between LNP membranes and cell membranes. Higher protein expression level. PEGylated Lipids Enhancing membrane fluidity DSPC – stabilizing lipid membrane structure, enhance nucleic acid encapsulation efficiency Targeting function extend circulation time immune responses (anti-PEG antibody) reducing clearance by blood proteins and macrophages reduce cellular uptake and hinder the escape of nanoparticles...

Generic Material Optimization siRNA materials DNA materials • Synthetic cap analogues and capping enzymes • Regulatory elements in the 5'-untranslated region (UTR) and the 3'-UTR • Poly(A) tail stabilizes mRNA and increases protein translation • Modified nucleosides, decrease innate immune activation and increase translation • Sequence and/or codon optimization increase translation o Increase metabolic stability and reduce degradation Phosphorothioate (PS) o Terminal backbone stabilization RISC loading, 5' phosphate modification o Prolonged durability on target silencing GNA glycol nucleic acid,...

High throughput system for LNP preparation Parallel multi-sample synthesis Microfluidic mixing Sequential single-sample synthesis Automatic high throughput system Parallel multi-sample synthesis Pipette mixing

High throughput system for LNP preparation PreciGenome NanoGenerator® Sequential microfluidic single- Flex-S Plus sample mixing Mixing Methods Microfluidic mixing Microfluidic mixing Pipette mixing Synthesis Mode Washing Needed Sample volume Sample conc. range Only low lipid concentration (1-2mM lipid) LNP size difference compared to scale up production LNP PDI compared to scale up production EE% compared to scale up production Protocol optimization Well developed Well developed Intense (ratio, speed, concentration, tip choice, etc.)

NanoGenerator® Flex-S Plus NanoGenerator ® Flex-S/Flex-S Plus Dead volume per sample Source of dead volume Typical production volume Minimum input volume (Aqueous :Lipid = 3:1) Estimated minimum mRNA cost Syringe Pump Systems Tubing Connection Systems Syringe, connector, Tubing, connector, mixing Chip and/or mixing chip NanoGenerator® Flex-S NanoGenerator® Flex-S Plus PreciGenome

• The Flex-S Plus System facilitates the rapid screening of nanoparticle formulations and early-stage payload candidates. • With a max throughput of 48 samples per run, 96 samples in 90 mins, the Flex-S Plus greatly streamlines screening processes. It offers comprehensive automation of complex protocols, enabling users to concentrate on other laboratory duties. • The system also permits experimentation with 100 -500 ul of samples while providing control over collection volumes. This allows users to optimize the use of valuable materials. • Library preparation function enables lipid/payload formulation...

NanoGenerator® Flex-S Plus Model Flex-S Plus Full automation Library preparation Buffer exchange Total flow rate Flow rate ratio Custom design flow rate Rapid screening of LNP formulations Rapid screening of payload Encapsulation efficiency Disposable consumables DNA, mRNA, siRNA, Protein, small mol ecules, etc. DNA, mRNA, siRNA, Protein, small mol ecules, etc. Optional functions: formulation library prep, buffer exchange, temperature control, HEPA filter, UV sterilization light Size range Payload HEPA filter/UV light Dimen

NanoGenerator® Flex-S Plus • High throughput workflow Change consumables Library prep. （optional） Change consumables Buffer exchange (optional) Final product • Automation workflow One instrument: offering two workflows • High throughput workflow Automation workflow (no need to change consumable) Raw materials Library prep. Buffer exchange Final produc

Screening reagents including: • Helper lipid • lonizablelcationic lipid • PEGylate lipid • Sterol lipid • Lipid combination Library preparation Cell studies with up to 32 formulations per run, 96 samples per hour Payload library • Payload Concentrations Carrier library • Lipid combination • Lipid ratio • Lipid Concentration Mixing condition • Total flow rate • Flow rate ratio Sample Workflow: 1. Load samples in 96 well plates; 2. Seal the 96 well plate (optional); 3. Put consumables on the deck: Chips, 96 well plates, pipette tips, and Gaskets; 4. Set parameters in the software and run the program;...

User friendly UI: 1. Up to 48 samples (12 chips) per run; 2. Different chips can apply different fluid parameters or/and product volumes 3. In one chip, up to 4 different formulations can be synthesized; 4. Offer recipe exporting and importing. Setting can be done in other computers; 5. Offer printing function. Layout can be printed out for sample prep guidance; 6. Clear consumable layout to help set up consumables. PrcciGcnomc PreciGenome Page 12

Flex-S Plus • Robust multi-sample synthesis • Reliable performance • Consistent results Model Flex-S Plus Aqueous phase Sodium acetate buffer, 100mM, pH5.2 Solvent phase LipidFlex, 15mM in ethanol

Example • 32 sample screening (formulation & N:P ratio screening) • 32 samples done in 30 mins Model Flex-S Plus 40 Aqueous phase RNA in Sodium acetate buffer, 100mM, pH5.2Solvent phase Different lipid formulation PrcciGcnomc PreciGenome Page 14

Library prep. • User friendly UI • Optional functions to help library prep. form raw materials; • Up to 12 lipid formulations (4 replicates) per run; • Automatically generated the volume of raw material need to be prepared; • Payload stock solution calculator helps prepare payload stocks based on different N/P ratio; Automatically generate LNP synthesis recipe based on the library prep setting. PrcciGcnomc PreciGenome Page 15