Antibiotics are a class of therapeutic molecules widely employed in both human and veterinary medicine. This article reviews the most recent advances in the analysis of antibiotics by CE in pharmaceutical, environmental, food, and biomedical fields. Emphasis is placed on the strategies to increase sensitivity as diverse off-line, in-line, and on-line preconcentration approaches and the use of different detection systems. The use of CE in the microchip format for the analysis of antibiotics is also reviewed in this article. Moreover, since the use of antibiotics as chiral selectors in CE has grown in the last years, a new section devoted to this aspect has been included. This review constitutes an update of previous published reviews and covers the literature published from June 2011 until June 2013.