This work describes the characterisation and culture performance of a novel, orbitally shaken, single-use photobioreactor (SUPBr) system for microalgae cultivation. The SUPBr mounted on an orbitally shaken platform was illuminated from below. Investigation of fluid hydrodynamics indicated a range of different flow regimes and the existence of ‘in-phase’ and ‘out-of-phase’ conditions. Quantification of the fluid mixing time (tm) indicated a decrease in tm values with increasing shaking frequency up to 90 rpm and then approximately constant tm values in the range 15–40 s. For batch cultivation of Chlorella sorokiniana, the highest biomass concentration achieved was 6.6 g L−1 at light intensity of 180 μmol m2 s−1. Doubling the total working volume resulted in 35–40% reduction in biomass yield while shaking frequency had little influence on culture kinetics and fatty methyl esters composition. Overall this work demonstrates the utility of the SUPBr for early stage development of algal cultivation processes.