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During World War I (WWI), infectious diseases including tetanus were among the most important causes of death. Even though its efficacy was somewhat controversial before the war, tetanus antiserum played a key role in reducing the mortality of this disease. A vial of tetanus antiserum dating back from WWI, left behind on the French battlefield by the US Army, was borrowed from a private collection and opened. The serum contained within was characterized by orthogonal biochemical techniques to determine if any neutralizing IgGs could remain after 100 years of storage. In vitro analysis by Size Exclusion Chromatography and Serum Protein Electrophoresis suggested the presence of residual IgG. In spite of our hopes, these IgGs were not able to protect mice against tetanus toxin challenge in a neutralizing assay. Even though our results indicate the presence of remaining IgGs inside the serum, they were functionally disabled. These results show that obscurity alone is insufficient to protect IgGs from degradation over very long periods of time at room temperature.    Highlights     Tetanus antiserum found its place in the therapeutic arsenal during World War I    A century-old vial of tetanus antiserum was opened for biochemical and in vivo characterization    Biochemical assays revealed the presence of proteins having all the characteristics of IgGs    The serum was unable to protect mice against toxinic challenge

History, extensive characterization and challenge of anti-tetanus serum from World War I: exciting remnants and deceived hopes