The orderly stacked laminate structures and extraordinary physicochemical properties of graphene oxide (GO) enable this material to serve as a nano-building block for separation membranes. However, attaining GO that has orderliness and regularity on three-dimensional substrates is still a great challenge. A new multiple tailored assembly is proposed to fabricate ethylenediamine-cross-linked GO composite membranes on a macroporous tubular ceramic substrate. X-ray diffraction, gas transmission rate and contact angle residence time analyses indicate that the GO nanosheets obtained from this method are neatly arranged, thus demonstrating the construction of efficient nanochannels for mass transfer. The resulting mul-EGO-60 membranes feature a permeability of 375.75 Lm(-2)h(-1)MPa(-1), which is 5.62 times as high as the pristine GO membranes without compromising the rejection (98.60%) of eriochrome black T. The flux and rejection for Na2SO4 are 237.72 Lm(-2)h(-1)MPa(-1) and 78.32%, respectively. The multiple tailored assembly offers an effective method for preparing high-performance two-dimensional GO membranes for mass separation.