Abstract Non-small cell lung cancer (NSCLC) remains the leading cause of cancer death in the United States despite a decline in mortality. Adenocarcinoma of the lung usually evolves from the mucosal glands and represents about 40% of all lung cancers. A significant proportion of lung adenocarcinoma patients present alterations in oncogenic driver genes. Next-Generation Sequencing (NGS) allows the sequencing of several genomic regions in a single test. There are multiple NGS approaches including Targeted Panel Sequencing (TPS), Whole Exome Sequencing (WES), and Whole Genome Sequencing (WGS). Methods: The detection of actionable genomic mutations in 60 lung adenocarcinoma surgical specimens was evaluated through two NGS approaches TPS and WGS. TPS was performed using the Seoul National University Hospital (SNUH) TPS consisting of 75 total genes, with a total coverage of 1.92Mb (exon 0.69Mb, intron 1.23Mb). WGS was performed using Illumina’s NovaSeq 6000 platform and data were processed with BWA and variant callers including Mutect2 and Delly. Functional mutations were determined by Ensembl VEP, while putative driver mutations were identified via the Genome Insight pipeline (San Diego). Results: A total of 120 driver mutations were detected by TPS. WGS identified 114 out of the 120 driver mutations (95% match). An additional 9 driver mutations were detected by WGS, along with thousands of additional non-driver mutations. (Table 1) Conclusion: An NGS-based approach allows for the identification of actionable molecular changes which leads to more informed clinical decision making. These results validate that Genome Insight’s WGS workflow and pipeline are comparable to TPS in detecting key driver mutations. Furthermore, this study demonstrated that WGS provides a more comprehensive genetic characterization of lung cancer, which may impact the therapeutic options and patients’ prognosis. Table 1. Driver mutations # of mutations detected by TPS # of mutations detected by WGS Detected by TPS EGFR 20 19 KRAS 9 9 TP53 30 30 Fusion oncogenes 17 17 PIK3 genes 1 1 BRAF 2 2 Secondary drivers detected by TPS 41 36 Sub-total 120 114 Missed by TPS Other drivers not detected by TPS 0 9 Total 120 123 Citation Format: Ji-Hyung Park, Young Tae Kim, Erin Connolly-Strong, Young Seok Ju. Whole genome sequencing provides a more comprehensive genetic characterization of lung cancer compared to a targeted panel sequencing [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 261.