Provided are a method of preparing an antibody with a regulated, sugar chain content, the method including the step of culturing antibody-expressing cells in a medium including glycerol as an additive for regulating the antibody sugar chain content, a method of preparing a high-quality population of antibodies by regulating the sugar content of the antibody to a desired content, and a population of antibodies prepared by the method. Further, provided is a method of regulating the antibody sugar chain content, the method including the step of culturing antibody-expressing cells in a medium including glycerol as an additive for regulating the antibody sugar chain content. Furthermore, provided is a medium composition, for regulating the antibody sugar chain content, the medium composition including glycerol as an additive for regulating the antibody sugar chain content. The preparation method of antibodies according to the present invention may be used to prepare a desired high-quality population of antibodies by regulating the sugar chain content of the antibody. Further, in terms of the development of biosimilars, the method of the present, invention may be used to regulate the sugar chain, content of antibodies, thereby preparing antibodies having high equivalence to a control drug. Since the sugar chain content may be regulated by a medium composition, the regulation method is easy and efficient in terms of time and cost, and therefore, widely applied to the fields of antibody preparation.

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