Analyses on diluted sera would not measure the complete activity of the natural in-vivo serum environment used by humoral immune responses. Since these humoral defences must react rapidly, serum reactions occurring during 10 and 60 s were analysed. Primo- and multiple-vaccinated, efficiently responsive, and protected animals were differentiated. A variation in the responses, particularly the quality of response, of individual animals was also now discernible. The critical importance of antibody-antigen contact time was demonstrated, wherein natural immunoglobulins and non-immunoglobulin opsonins would influence specific antibody reactivity, through an increased avidity of reaction with antigen. Although not measured directly, the influence of the non-specific serum components would be manifest through increased specific antibody binding. By considering serum as an entity, analysis of all constituent components permits increased qualitative assessment of post-vaccination sera.