This article attempts to provide an overview of different approaches proposed to improve the performance of immunoassay techniques, most of which are used in routine clinical analysis and are finding wide acceptance in other analytical areas, such as environmental and food analysis. The term
immunoassay
here encompasses all the methods that involve the formation of antigen‐antibody complex to attain the separation and/or detection of analytes, including conventional immunoassays, immunosensors, and immunoaffinity techniques. Advances in the production of antibodies, in the availability of new labels, and in the immobilization of immunoreagents, which are allowing the improvement of the selectivity, sensitivity and robustness of these assays, are described. The recent use of different nanomaterials as labels, nanoscaffolds, and separation media in immunoassay techniques is especially emphasized. Also, the development of new approaches to increase the usefulness of immunoassays for multianalyte detection is shown through the different microarray systems proposed. The efforts carried out to miniaturize immunoassays are described by discussing the usefulness of microfluidic systems. Finally, recent achievements in immunoextraction, alone or in combination with other separation techniques, such as liquid chromatography and capillary electrophoresis, are also included.