Aqueous Two-Phase Systems (ATPS) is a primary recovery technique that has shown great potential for the efficient extraction and purification of high value biological compounds. The main advantages of this technique include scaling up feasibility, process integration capability and biocompatibility. In this review, the efficient use of ATPS for the extraction of proteins, genetic material, low molecular weight compounds, bioparticles, nanoparticles and cells is highlighted. The important role of ATPS in process integration, i.e., extractive conversion, extractive fermentation, cell disruption integrated with product recovery, and extractive purification, is discussed. A novel approach to protein molecular characterization combining ATPS and 2-dimension electrophoresis (2-DE) is introduced as a first step in the process development. Novel approaches for downstream processing using ATPS and dielectrophoresis are presented. Finally, trends concerning the application of ATPS strategies to address the future challenges of bioseparation are discussed.

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Antibodies for therapeutic use are being continuously approved and their demand has been steadily growing. As known, the golden standard for monoclonal antibody (mAb) purification is Protein A affinity chromatography, a technology that has gained high interest because of its great performance and capabilities. The main concerns are the elevated resins costs and their limited lifetime compared to other resins (e.g. ion exchange chromatography). Great efforts have been carried out to improve purification conditions, such as resin characterization and designing alkali/acid stable resins with a longer lifetime. Modification of Protein A ligands and alternative formats such as monoliths membranes and microshperes have been tested to increase the purification performance. New technology has been proposed to improve the large‐scale separation; in addition, alternative ligands have been suggested to capture mAbs instead of Protein A ligand; however, most of the information is locked by pharmaceutical companies. This mini review summarizes and describes the advances, results, and impact on the Protein A chromatography purification processing.

Electrochemical immunosensors, EIs, are systems that combine the analytical power of electrochemical techniques and the high selectivity and specificity of antibodies in a solid phase immunoassay for target analyte. In EIs, the most used transducer platforms are screen printed electrodes, SPEs. Some characteristics of EIs are their low cost, portability for point of care testing (POCT) applications, high specificity and selectivity to the target molecule, low sample and reagent consumption and easy to use. Despite all these attractive features, still exist one to cover and it is the enhancement of the sensitivity of the EIs. In this review, an approach to understand how this can be achieved is presented. First, it is necessary to comprise thoroughly all the complex phenomena that happen simultaneously in the protein-surface interface when adsorption of the protein occurs. Physicochemical properties of the protein and the surface as well as the adsorption phenomena influence the sensitivity of the EIs. From this point, some strategies to suppress non-specific binding, NSB, of proteins onto electrode surfaces in order to improve the sensitivity of EIs are mentioned.

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