The great success of electrogenerated chemiluminescence (ECL) in analytical chemistry can be measured by the widespread use of the technology in different fields, ranging from basic research to commercial clinical and biological applications. Indeed, this remarkable readout method offers intrinsic advantages by comparison to other transduction methods: high sensitivity, extremely wide dynamic range and insensitivity to matrix effects. In addition, its versatility allows exploiting various types of biomolecular interactions and therefore to detect specifically targeted analytes of biological interest such as proteins, nucleic acids and enzymatic substrates. Numerous assay formats, biosensors or analytical strategies with new ECL labels or with label-free approaches have been proposed by using nanostructured materials: carbon nanotubes, metal or doped nanoparticles, graphene, carbon dots, quantum dots or ultrathin films. The development of analytical ECL has also been fueled by discovering novel luminophores and efficient coreactants and also by deciphering the complexity of the ECL mechanisms at the minute scale. The combination of ECL with microfluidics, paper-based materials, bipolar electrochemistry, and portable miniaturized devices has led to various intriguing and promising analytical applications.