Recent developments in three-dimensional printing (3DP) have attracted the attention of analytical scientists interested in fabricating 3D devices having promising geometric functions to achieve desirable analytical performance. To break through the barrier of limited availability of 3DP materials and to extend the chemical reactivity and functionalities of devices manufactured using conventional 3DP, new approaches are being developed for the functionalization of 3D-printed devices for chemical and biochemical analysis. This Review discusses recent advances in the chemical functionalization schemes used in the main 3DP technologies, including (i) post-printing modification and surface immobilization of reactive substances on printed materials, (ii) pre-printing incorporation of reactive substances into raw printing materials, and (iii) combinations of both strategies, and their effects on the selectivity and/or sensitivity of related analytical methods. In addition, the state of the art of 3D-printed functionalized analytical devices for enzymatic derivatization and sensing, electrochemical sensing, and sample pretreatment applications are also reviewed, highlighting the importance of introducing new functional and functionalized materials to facilitate future 3DP-enabled manufacturing of multifunctional analytical devices. Display omitted •New approaches are being developed to chemically functionalize 3D-printed devices.•Major schemes are post-printing immobilization and pre-printing incorporation.•D-printed devices functionalized for chemical and biochemical analysis are included.•Enzymatic sensing, electrochemical sensing, and sample pretreatment are reviewed.