Company: BLLN
Filing Date: 2025-08-11
Form Type: DRS/A
Source: 0000950123-25-007483
Chunk: 203

Company: BillionToOne, Inc.
Filing Date: 2025-08-11
Form: DRS/A
Chunk 203
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 of less than 10% for a typical sample with 1% tumor fraction, which is at least two times lower than conventional tumor-naive, targeted-panel methods that measure VAF. It has demonstrated a strong correlation with clinical outcomes in patients with lung, colorectal, and pancreatic cancers.

| 59 |     | Yu P, Vlachogiannis G, Snyder MW, et al. Molecular counting enables accurate and precise quantification of methylated circulating tumor DNA for tumor-naive cancer therapy response monitoring. Sci Rep. 2025;15:5869 |

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1000 Tumor Methylation Score 0 750 500 250 Breast Analytical LOD is at 0.01% Colorectal 0% 0.1% 0.2% 0.3% 0.4% 0.5% 900 600 300 0 0% 0.1% 0.2% 0.3% 0.4% 0.5% Contrived Tumor Fraction

Northstar Response assay measures changes in tumor burden across 12+ cancer types Brain Breast Colorectal Endometrial Liver Lung Ovarian Pancreas Prostate Renal Sarcoma Stomach

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We also collaborated with University of California, San Diego on a study involving immunotherapy and immuno-chemotherapy treated advanced non-smallcell lung cancer patients. 60In that study, which involved 60 samples from 51 patients, Northstar Response’s changes in TMS measured four to ten weeks after starting treatment significantly predicted real-world progression free survival (rwPFS, p < 0.0001), compared to standard imaging assessments which did not reach statistical significance (p = 0.55). The p-valueis used to determine the probability as to whether the difference between two data sets is due to chance. The smaller the p-value,the more likely the differences are not due to chance alone. In general, if the p-valueis less than or equal to 0.05, the outcome is considered statistically significant. The study also showed that the test often detected treatment response and progression earlier than standard of care CT scans, with high concordance between TMS and clinical outcomes. 100 RECIST Objective Response Probability of Survival 0 50 75 25 Partial Response Stable Disease Progressive Disease P:0.55 0 100 200 300 Real-World Progression-Free Survival (Days) 400 Probability of Survival