This study aimed to investigate the effect of housing system and dietary biotin supplementation on productive performance, blood biochemical parametersand economic efficiency of Benha line chicken. A total of 224 chickens, 20 weeks old with similar body weight were used in this study, birds were equally divided into two main groups in a factorial arrangement design (2x4x3). Pullets of the first group reared in cages. While, pullets of the second group reared on deep litter. The pullets of each group were subdivided into four sub-groups (each of 25 females and 3 males) according to dietary biotin supplementation. Birds of the 1^st sub-group were fed on basal layer diet and considered as control, while the 2^nd, 3^rd and 4^th sub-groups fed basal diet supplemented with biotin at levels of 100, 150 and 200µg /kg feed, respectively.  The results revealed that, pullets reared in cages significantly improve body weight, feed conversion, early to reach age at sexual maturity, egg production rate, egg weight, egg mass, absolute yolk weight compared to those reared on deep litter. While, pullets reared on deep litter had significantly improved feed consumption, egg shell thickness, plasma calcium, inorganic phosphorus and decreased plasma cholesterol and triglycerides compared to those reared in cages. Biotin supplementation at a level of 100 µg /kg feed had significantly increased body weight, egg weight, absolute and relative egg albumen weight, plasma inorganic phosphorus and decreased feed consumption compared to the other levels. While, pullets fed on diet supplemented 150µg biotin /kg feed showed the improving in feed conversion, early sexual maturity, increased egg production, egg mass and plasma calcium compared with the other levels. In addition, birds fed diet supplemented with biotin at a level of 150 µg/kg feed and reared in cages recorded the highest economic efficiency (219.04%), followed by those fed diet supplemented with biotin at a level of 100 µg/ kg feed and reared on deep litter (141.37 %) compared to the other levels and control group. From the economic point of view, it could be concluded that rearing layer chickens in cages and fed on diet supplemented with biotin at a level of 150µg/kg feed seemed to be adequate to achieve the favorable results, followed by those reared on deep litter and fed diet supplemented with biotin at a level of 100 µg/ kg diet.