Tuberculosis remains a leading global cause of morbidity and mortality. While the World Health Organization has established target product profiles to guide diagnostic development, descriptions of potential use cases for biomarkers and tests remain scattered across diverse guidelines and literature. There is currently no clear synthesis or prioritization of these use cases to guide research and investment. This protocol describes a study aiming to identify, map, and prioritize tuberculosis biomarker use cases to focus development efforts where they can maximize clinical and public health impact. Methods The study will employ a multi-step, iterative process combining a scoping review with a consensus-building exercise. First, a scoping review following the Arksey and O’Malley framework will be conducted across academic databases and global health repositories to identify existing use cases. Identified use cases will be mapped to a framework of tuberculosis stages, including infection, disease, treatment, and post-tuberculosis care. Subsequently, a hybrid RAND/UCLA appropriateness method and modified Delphi process will be used to establish consensus among a diverse group of stakeholders, including researchers, clinicians, industry representatives, and people with lived experience of tuberculosis. Stakeholders will participate in anonymous online surveys to rate the validity and importance of each use case, followed by hybrid in-person and virtual meetings to resolve areas of disagreement. Results The primary outputs will be a consolidated, prioritized list of tuberculosis biomarker use cases. These will be disseminated through a public report, peer-reviewed scientific articles, and conference presentations to inform the global research community and funders. Conclusions By integrating evidence synthesis with structured stakeholder consensus, this project seeks to align biomarker discovery and diagnostic development with the most critical clinical and public health needs, ultimately reducing research waste and supporting the global target of ending the tuberculosis epidemic.