Application of longitudinally profiled (LP) steel plate in the flange of flexural members may provide a good solution to optimize their mechanical performance and to improve the efficiency of steel use, whilst existing design codes provide no design guidance or prediction methodology for such advanced beams in terms of flexural behaviour. To clarify their flexural strength and rotation capacity, tests on two full-scale welded I-section steel beams with longitudinally profiled flanges (LPB members) are carried out herein, as well as two traditional beams with uniform cross-section for comparison. All the specimens exhibit sufficient flexural strength and rotation capacity for seismic plastic design, and specifically, the LPB members possess even better performance in case of identical steel usage. Parametric analyses of 250 beams incorporating a wide range of flange slenderness and steel grades, are conducted by employing the validated nonlinear FE model to investigate the effects of rate of thickness change for the LP flanges. The results show that the effect is limited on the flexural strength but significant on rotation capacity. The existing design provisions for beams with uniform cross-section give generally conservative design results for the flexural strength of the LPB members, but limiting values of flange slenderness needs to be reduced. The research outcomes may provide an important basis for promoting the application of LP plates in flexural members.