In biomedical research and diagnostic medicine, as well as in cellular therapy procedures, cell separation has become crucial. The process enables the isolation of specific cell populations, such as cells with a specific antigen, from a mixture of different cells. One such antigen or cell marker is the CD3 molecule. Over the past two decades, this process has undergone remarkable development, leading to various methods for isolating specific cell types. In our study, we focused on analyzing and comparing three methods of cell separation: PluriBead®, Dynabeads, and Strep-tag®. We initially used the PluriBead® and Strep-tag® methods to separate CD3+ (Jurkat) and CD3- (Kasumi-1) cell lines. With all three methods, we separated primary cells from peripheral blood obtained through leukapheresis. The aim was to evaluate the efficiency, specificity, and economic justification of all three methods for separating target cell populations. We focused on separating cells with antibodies against CD3. The Strep-tag® method achieved a high yield with the CD3+ Jurkat cell line, while PluriBead® achieved a lower yield, indicating greater effectiveness of the Strep-tag® method in separating these cells. For the CD3- Kasumi-1 cell line, both systems (Strep-tag® and PluriBead®) achieved equally low isolation results, as expected, since Kasumi-1 cells do not express the CD3 antigen to a large extent, meaning the separation was appropriately negative. Dynabeads proved to be an effective compromise for separating leukocyte concentrate, as we achieved moderate yield with high purity and acceptable viability using this method. However, the Strep-tag® and PluriBead® methods were superior in certain characteristics. With Strep-tag®, we achieved higher yield with similar purity, while with PluriBead®, we obtained better viability despite lower yield. From an economic standpoint, the Dynabeads method required the highest initial investment, while the PluriBead® method had the lowest recurring costs, and Strep-tag® had high repetition costs but no initial investment. Our findings provide important insights into cell separation methods. Although each method has its unique advantages, their selection depends on the specific needs and limitations of the project.
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