This paper reports a fundamental study of lubricant film formation with model synovial fluid components (proteins) and bovine serum (BS). The objective was to investigate the role of proteins in the lubrication process. Film thickness was measured by optical interferometry in a ball-on-disc device (mean speed range of 2–60 mm/s). A commercial cobalt–chromium (CoCrMo) metal femoral head was used as the stationary component. The results for BS showed complex time-dependent behaviour, which was not representative of a simple fluid. After a few minutes sliding BS formed a thin adherent film of 10–20 nm, which was attributed to protein absorbance at the surface. This layer was augmented by a hydrodynamic film, which often increased at slow speeds. At the end of the test deposited surface layers of 20–50 nm were measured. Imaging of the contact showed that at slow speeds an apparent ‘phase boundary’ formed in the inlet just in front of the Hertzian zone. This was associated with the formation of a reservoir of high-viscosity material that periodically moved through the contact forming a much thicker film. The study shows that proteins play an important role in the film-forming process and current lubrication models do not capture these mechanisms.