BACKGROUND T-lymphocyte-based immunotherapy has been widely studied and applied in experiments and clinics. T lymphocytes need to be enriched and amplified from peripheral blood in T lymphocyte immunotherapy. However, enrichment and expansion of T lymphocytes are performed using separated procedures in popular protocols, leading to tedious steps as well as increasing the possibility of cell contamination during clinical application. Herein, we prepared artificial antigen-presenting immunomagnetic beads that can be used for both enrichment and expansion of T lymphocytes from peripheral blood, which could greatly simplify the procedures of T lymphocyte immunotherapy. Anti-CD3 and anti-CD28 monoclonal antibodies were crosslinked to poly(acrylic acid) magnetic microspheres of about 300 nm in diameter to make artificial antigen-presenting immunomagnetic beads that combine the separation and activation functions of T lymphocytes. RESULTS The experimental results showed that the prepared beads could both enrich T lymphocytes from peripheral blood with about 80% enrichment efficiency and also successfully activate T lymphocytes in culture. In vitro cell experiments demonstrated that the activated T lymphocytes expanded using the immunomagnetic beads could kill cells of tumor cell line K562. CONCLUSIONS The results demonstrate that the immunomagnetic beads prepared in our study can realize the enrichment and expansion of T lymphocytes from peripheral blood mononuclear cells, and provide a convenient tool for the integrated design of clinical immunotherapy instruments. (c) 2020 Society of Chemical Industry