Abstract Background Hepatitis B is a liver infection caused by the Hepatitis B virus. While preventable by vaccination, Hepatitis B remains prevalent across the globe. While prevalence is low in many areas of the world, there are still regions in which the prevalence is considered to be high. Hepatitis B surface antigen (HBsAg) can be one of the first indicators of Hepatitis B infection, and it is a commonly screened diagnostic test to determine if a patient is immune, susceptible, or has acute or chronic HBV infection. While HBsAg is a critical marker in diagnosing a patient, it presents unique challenges. The Thermo Scientific™ MAS™ Omni™ Infectious BSI Positive Control Panel* is a multi-constituent IVD control containing anti-HIV-1/2, anti-HBc, anti-HCV, and anti-HTLV I/II antibodies, as well as Hepatitis B surface antigen (HBsAg) in a human plasma-based matrix. Development and Stability Studies were conducted to evaluate and ensure optimal analyte performance throughout the shelf-life duration. As this is a multi-constituent control, the HBsAg challenges must be adequately addressed to ensure the integrity and reliability of the results of all analytes. Two specific areas of focus are HBsAg instability at increased temperature and HBsAg-Fibrinogen interaction in plasma products. We report here the continued development and assessment of HBsAg in the MAS™ Omni Infectious Controls family. Methods The BSI Positive Control Panel was evaluated with IVD assays to assess Shelf-Life and demonstrate the effect of elevated temperature and fibrinogen on HBsAg. Shelf life of the products was determined by real-time, accelerated, open vial, and in-use stability monitoring. Additionally, open-vial stability monitoring demonstrates the relationship between HBsAg and fibrinogen within the control products. Between Month 0 and 12, samples were run neat without processing. At Month 12, samples were centrifuged to verify the validity of the open-vial stability claim. Results The evaluation of the BSI Positive Control Panel demonstrates the effects of temperature and fibrinogen on HBsAg. The Real-Time and Accelerated stability studies combine to demonstrate the effect of elevated temperatures on HBsAg within the multi-analyte control. The Open-Vial stability studies completed at 0 and 12 months combine to demonstrate the effect of fibrinogen on HBsAg within the multi-analyte control. Conclusions The MAS™ Omni Infectious control products will perform optimally provided the proper sample processing and storage conditions are implemented. Improper storage at elevated temperatures will result in degradation at an increased rate for the HBsAg present in the sample. Additionally, it is imperative that the proper raw materials are chosen in the production of the multi-analyte controls. When using plasma products, it is best to use defibrinated plasma. For extra precaution, raw material should be centrifuged and filtered prior to production to avoid degradation due to fibrinogen. Reliable, high-quality controls are vital in ensuring that clinical diagnostic assays are functioning properly. *Availability of product in each country depends on local regulatory marketing authorization status