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Adoptive cell transfer represents an important mode of cancer immunotherapy and posttransplant associated viral infections. This technique involves massive volume reduction, as the procedure starts with large-scale ex vivo expansion of lymphocyte cultures (volume up to 60 L), which are harvested at the end of the in vitro process and terminates in a small volume to be infused into the patient. Toward this end, we develop a novel efficient process based on the COBE Spectra apheresis machine usually used for apheresis process. As the COBE Spectra cell separator is easy to use and often available at medical centers, this novel technique is applicable to many transplant and cell processing centers. Our results show a high recovery yield (98%±15%) and viability (ranged 79% to 99%) of the large-scale expanded lymphocytes. It preserves sterility of the product and is therefore suitable for immunotherapy treatments.