The slide is titled "NR0754 Run Control" and serves as a title slide for a presentation. Its subtitle summarizes the production issue and resolution status as of December 2025.

NR0754 Run Control

Production Issue Summary & Resolution Status – December 2025

The agenda slide outlines a presentation on a production issue, starting with an introduction to objectives, scope, timeline, involved teams, and NR0754. It proceeds to summarize the issue investigation, root cause analysis with Pippin purification findings and fixes, and concludes with results like TVER reduction, next steps, stability testing, and key takeaways.

Agenda

1. Introduction and Background

Objectives, scope, timeline, teams involved, and NR0754 introduction.

2. Issue Summary and Investigation

Production issues, data comparisons, charts, and tested factors summary.

3. Root Cause and Corrective Actions

Pippin purification findings, experimental data, and implemented fixes.

4. Results, Next Steps, and Takeaways

Post-fix TVER reduction, stability testing, and lessons learned. Source: NR0754 Run Control – Production Issue Summary & Resolution Status (Dec 2025)

NR0754 was launched in July 2025 as a replacement for NR0339, designed to closely mimic patient sample preparation while incorporating a Pippin purification step and using Quant-it for precise quantification. This ensures consistent performance in run controls.

Background

• NR0754 launched July 2025 as NR0339 replacement
• Designed to mimic patient sample preparation
• Incorporates Pippin purification step
• Uses Quant-it for accurate quantification
• Ensures consistency in run control performance

The slide summarizes a production issue involving elevated TVER levels in run controls for batch NR0754, impacting sites in Austin CLIA, San Carlos CLIA, and Austin CTA, with the increase roughly doubling the baseline from NR0339. Importantly, the issue is limited to production run controls and does not affect patient sample processing.

Production Issue Summary

• Elevated TVER observed in NR0754 run controls
• Impacted sites: Austin CLIA, San Carlos CLIA, Austin CTA
• No adverse effects on patient sample processing
• TVER increase approximately 2x over NR0339 baseline
• Issue confined to production run controls only

The Data Summary Chart highlights a 2x increase in TVER for NR0754 compared to the NR0339 baseline. It notes elevated transversions across 3 affected sites, while transition rates remain stable with no significant changes observed.

Data Summary Chart

• 2x: TVER Increase

NR0754 vs NR0339 baseline

• 3: Affected Sites

Elevated transversions across all

• Stable: Transition Rates

No significant change observed Source: Internal Production Data

The slide summarizes an investigation into elevated Total Transversion Error Rate (TVER), ruling out factors like sequencers, reagents, lots, and cell source after extensive testing. It suspects the formulation process, especially the Pippin purification step, as the cause due to the introduction of reactive oxygen species.

Investigation Summary

The slide titled "Root Cause Findings" highlights key issues in a process, noting that Pippin purification introduces reactive oxygen species, which elevate TVER levels when exposed to light afterward. It concludes that buffer exchange effectively mitigates this TVER elevation.

Root Cause Findings

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• Pippin purification introduces reactive oxygen species.
• Post-Pippin light exposure raises TVER levels.
• Buffer exchange mitigates TVER elevation.

Source: Reactive oxygen species

The slide presents key experimental statistics supporting the research, highlighting a 2.5x increase in TVER under light exposure compared to dark conditions. It also covers 90% recovery efficiency via buffer exchange, 4.1% variation in operator consistency, and a minimum TVER of 1.5E-05 at 4°C for optimal temperature effects.

Supporting Experimental Data

• 2.5x: Light Exposure Effect

TVER increase in light vs dark study

• 90%: BEX Recovery Efficiency

Mitigation via buffer exchange

• 4.1%: Operator Consistency Variation

Low deviation across operators

• 1.5E-05: Minimum TVER at 4°C

Optimal temperature effect

To address issues in the purification process, corrective actions included adding a buffer exchange step after Pippin purification and introducing dark handling protocols to reduce light exposure. Additionally, operators were retrained on standardized procedures, and ongoing stability monitoring for lots was established.

Corrective Actions Implemented

• Implemented buffer exchange step post-Pippin purification
• Introduced dark handling protocols to minimize light exposure
• Retrained operators on standardized handling procedures
• Established ongoing monitoring of lots for stability

The slide presents pre- and post-fix Total Verification Error Rates (TVER), showing a baseline of 3E-05 reduced to 2E-05 after implementation, achieving a 33% improvement. It also notes that three specific lots—244005, 245072, and 246924—were evaluated in this analysis.

Results Post-Fix Chart

• 3E-05: Pre-Fix TVER

Baseline error rate

• 2E-05: Post-Fix TVER

Improved error rate

• 33%: TVER Reduction

Decrease after fixes

• 3: Lots Evaluated

244005, 245072, 246924

The "Next Steps" slide outlines key actions for the revised NR0754 formulation, including conducting stability testing, validating dye impact on transversion error rates, and implementing buffer exchange and dark handling protocols. It also targets CAPA closure by the end of Q1 2026 while monitoring production lots for ongoing TVER stability.

Next Steps

• Conduct stability testing on revised NR0754 formulation.
• Validate dye impact on transversion error rates.
• Implement buffer exchange and dark handling protocols.
• Target CAPA closure by end of Q1 2026.
• Monitor production lots for ongoing TVER stability.

Reactive oxygen species (ROS) from Pippin purification and light sensitivity are primary causes of elevated TVER, but corrective measures like buffer exchange, dark handling, and retraining have restored production stability. Ongoing monitoring of lots is crucial for quality, with a call to reach out for updates on validation and CAPA progress.

Key Takeaways

• Reactive oxygen species (ROS) from Pippin purification and light sensitivity are key contributors to elevated TVER.

• Implemented corrective actions (buffer exchange, dark handling, retraining) have restored performance stability.
• Ongoing monitoring of production lots is essential for sustained quality.

Closing Message: Production stability achieved through targeted fixes. Call-to-Action: Reach out for updates on validation and CAPA progress.