The single step-up pressure filtration test was developed to determine the pressure dependence of average specific resistance of the cake formed in ultrafiltration of a variety of nano-colloids over a wide range of pressure drops across the cake. The values of the average specific resistance at extremely low pressures were obtained from only the flux decline data through the use of the distinct time variation of the pressure drop across the cake generated by using the ultrafiltration membrane with a high hydraulic resistance under the low filtration pressure in the first step of filtration. The values at higher pressures were obtained from the time variation of the filtration rate induced by a stepwise increase in the pressure. The correlations between the average specific cake resistance and the pressure drop across the cake were evaluated using only the flux decline data for a variety of different proteins and nanoparticles.