Company: BLLN
Filing Date: 2025-08-11
Form Type: DRS/A
Source: 0000950123-25-007483
Chunk: 207

Company: BillionToOne, Inc.
Filing Date: 2025-08-11
Form: DRS/A
Chunk 207
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 D., Beduschi, T., Sahin, I., Hughes, S., Zarrinpar, A.          
 Link*: https://doi.org/10.1097/XCS.0000000000000939             
 Publication Date: April 2024                                    |     | This collaboration between BillionToOne and the University                                                                                                                                                                                              
 of Florida demonstrates the application of methylated circulating tumor DNA (ctDNA) quantification for monitoring hepatocellular carcinoma (HCC) treatment efficacy using BillionToOne’s proprietary molecular counting technology. Blood samples       
 from 25 patients (21 with HCC and 4 with benign liver masses) were collected at various treatment points to noninvasively assess tumor burden changes. Results confirmed that methylated ctDNA quantification reliably detected HCC and accurately      
 differentiated cancer patients from non-cancer controls. Compared to alpha-fetoprotein (AFP), the current standard biomarker, methylated ctDNA quantification showed significantly superior sensitivity (80.9% vs. 51.7%) and 100% specificity,         
 effectively detecting tumor presence and subtle changes in tumor burden post- transplantation, post-surgical removal, and throughout locoregional and systemic therapies. Furthermore, changes in the Tumor Methylation Score (TMS) demonstrated better 
 predictive                                                                                                                                                                                                                                              |

155

| accuracy for tumor progression than AFP alone (AUC=0.800 vs. AUC=0.783), and when combined with AFP,                                                                                                  
 produced the strongest predictive model (AUC=0.892). These findings strongly support the clinical utility of TMS as a powerful tool for guiding personalized therapeutic decisions in HCC management. |

Patient Case Studies We believe the following patient case studies are examples of how our patients can benefit from our tests, and highlight one of the important reasons behind physicians’ choice of utilizing our products and platform. We have highlighted patients across our UNITY and Northstar tests. Because these patients cover varied populations and circumstances, we believe the following case studies provide a helpful overview of the results that could be achieved by the broader patient population during the same time periods presented. However, patients experience different results depending on a number of factors, and these case studies are not necessarily representative of the results achieved by other patients. In particular, in a screening setting, such as for our prenatal tests, more than 99% of patients receive negative or low-risk results, which reduce the anxiety and the fear of the unknown for these patients, but do not necessarily improve their healthcare outcomes. UNITY Fetal Risk Screen