Lentiviral vectors (LVVs) are broadly used in research and therapeutic development, yet substantial interlaboratory variability continues to hinder the comparison of vector quantity, potency, and impurity measurements, underscoring the need for standardized reference materials. To address this need, the Lentiviral Vector Reference Material (LVVRM) initiative was launched as a structured, multi-phase effort, with earlier phases focused on large-scale upstream production and downstream preparation of a GFP-encoding LVV. In the final phase of the LVVRM initiative described herein, bulk material produced during prior large-scale manufacturing runs was thawed, pooled, subjected to sterile filtration, and dispensed into single-use vials under controlled conditions designed to maintain stability and consistency. Each lot then underwent comprehensive quality control testing. Sterility, identity, purity, physical and functional titers, and safety attributes were assessed using established characterization frameworks appropriate for lentiviral vector reference materials. The resulting LVVRM (ATCC® VR-3382™) met quality standards and release criteria, providing a well-characterized reagent suitable for assay calibration, method qualification, and longitudinal performance monitoring across laboratories.