Two field experiments were carried out at the experimental station farm Faculty of Agriculture (Saba –Basha), Alexandria University, Egypt, during 2013/2014 and 2014/2015 seasons to study the response of some wheat varieties (Triticum  aestivum, L.) (Masr-1, Sakha-94 and Gemmeiza-9) and study the effect of nitrogen levels (zero, 35, 70 unit nitrogen) and bio-fertilization (Control, 35 Kg/fed + Microbein, 75 Kg/fed + Microbein) on yield and technological characteristics of wheat using spilt plot design with three replicates. The preceding summer crop was cotton in both growing seasons. The results could be summarized as follows Wheat cultivars, i.e. (Sakha-94, Masr-1 and Gemmeiza-9) were significantly differed in all growth characters under study. Nitrogen levels and Bio-fertilizers produced significant increments in all growth characters and Chemical composition. The interaction between wheat cultivars, nitrogen levels and bio-fertilizers showed significant effects on all studied traits. The obtained results showed the highest values plant height, number of tillers m^-2, spike length, number of spike m^2, number of spikelets/spikes, number of kernels/ spikes 1000-grain weight, spike weight, Ardabs, grain, straw and biological yields, harvest index, wheat moisture %, ash%, falling number sec, specific weight sec, total protein%, wet gluten%, dry gluten%, gluten index %, flour moisture%, ash moisture%,  flour ash%, bran ash%, flour falling number sec, flour wet gluten% and milling characters%. Were obtained from the highest level of nitrogen 75 KG/fed N and 75 Kg/fed N+ microbein.