Cancer treatment not only damages tumour cells but also changes them. And some of these changes could be used to target treatment more effectively. Scientists from CEITEC Masaryk University and University Hospital Brno have found that during treatment for chronic lymphocytic leukaemia, a receptor becomes more abundant on the surface of cancer cells, making it possible to target them with an antibody. In laboratory experiments, the researchers therefore combined a commonly used drug with an antibody targeting this receptor. The combination killed leukaemia cells significantly more effectively than the drugs alone. The research is still at the preclinical stage, but points to a possible way towards more effective combination cancer therapies.Chronic lymphocytic leukaemia (CLL) is the most common leukaemia in older adults in the Western world. It affects white blood cells called B lymphocytes, which begin to multiply uncontrollably and accumulate in the blood, bone marrow and lymph nodes. CLL treatment has changed significantly in recent years. Modern targeted drugs can block signals that cancer cells need to survive and multiply.The researchers focused on what happens to leukaemia cells during such treatment. They analysed patient samples collected before and during treatment with BCR inhibitors, drugs commonly used to treat CLL. They studied patients receiving two different targeted BCR inhibitors. Although the two drugs affect cancer cells differently, the cells responded to both with surprisingly similar changes.<img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/CEITEC_Leukemicke_bunky_reaguji_na_lecbu_tak_ze_ze_sebe_udelaji_terc_2_8b86014944.jpeg" alt="CEITEC/Miroslav Boudný: Leukaemia Cells Respond to Treatment by Turning Themselves into a Target: Preparation of cells for experiments." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_CEITEC_Leukemicke_bunky_reaguji_na_lecbu_tak_ze_ze_sebe_udelaji_terc_2_8b86014944.jpeg 235w," sizes="100vw" width="1600" height="1067">“Cancer cells are not passive. They respond and adapt to treatment. We wanted to know whether, among the changes triggered by treatment, we could find something that could be used against them. That is what led us to the CXCR4 receptor,” explains <a target="_blank" rel="noopener noreferrer" href="https://www.ceitec.eu/veronika-sandova-ph-d/u95494?page=publication">Veronika Šandová</a>, a researcher&nbsp;from Marek Mraz’s laboratory at CEITEC Masaryk University and University Hospital Brno.CXCR4 is a receptor on the cell surface that, among other functions, helps leukaemia cells move to environments that provide favourable conditions for their survival. The researchers found that once treatment begins, CXCR4 levels on the surface of cancer cells increase significantly. The way cancer cells respond to treatment could therefore also become a weakness. Molecules on the cell surface are readily accessible to targeted treatment using specific antibodies.In laboratory experiments, the researchers therefore added ulocuplumab, an antibody that binds to the CXCR4 receptor, to the drugs already being used. The effect was substantial. While around seven out of ten leukaemia cells survived treatment with the drugs alone, only around three to four out of ten survived when the drugs were combined with the antibody. “It is a little like a cancer cell adapting against the first attack and, in doing so, marking itself as a target for a second one. The treatment increases the amount of a particular molecule on its surface, which we can then target,” Veronika Šandová explains.<img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/CEITEC_Leukemicke_bunky_reaguji_na_lecbu_tak_ze_ze_sebe_udelaji_terc_3_a3d50f3f82.jpeg" alt="CEITEC/Miroslav Boudný: Leukaemia Cells Respond to Treatment by Turning Themselves into a Target: Cooling the cells after the experiment prevents degradation of protein and other molecules." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_CEITEC_Leukemicke_bunky_reaguji_na_lecbu_tak_ze_ze_sebe_udelaji_terc_3_a3d50f3f82.jpeg 235w," sizes="100vw" width="1600" height="1067">The researchers went on to verify the results in a mouse model using leukaemia cells from patients. Here, too, the antibody targeting CXCR4 was highly effective at eliminating the cancer cells.The research is still at the preclinical stage, so this is not a new treatment that can yet be offered to patients. However, it suggests a possible way to identify more effective combinations of cancer drugs. “With cancer, targeting a single mechanism is often not enough because cancer cells can respond to treatment and find other ways to escape and survive. Our results show that we can use these changes to our advantage and target them with additional treatment,” says <a target="_blank" rel="noopener noreferrer" href="https://www.ceitec.eu/prof-marek-mraz-ph-d/u26692">Marek Mráz</a>, head of the research laboratory where the study was conducted.The study therefore offers another perspective on the search for new combinations of cancer treatments: looking not only at whether a drug kills a cancer cell, but also at how it changes the cell – and whether any of these changes could provide a new target for further treatment.The study was conducted in collaboration among scientists from CEITEC Masaryk University, University Hospital Brno, the Faculty of Medicine at Masaryk University, and the Dana-Farber Cancer Institute in Boston.The results were published in the scientific journal <a target="_blank" rel="noopener noreferrer" href="https://onlinelibrary.wiley.com/doi/10.1002/hem3.70481">HemaSphere</a>.<blockquote><h4>The original article</h4><h4><a target="_blank" rel="noopener noreferrer" href="https://onlinelibrary.wiley.com/doi/10.1002/hem3.70481">Gene expression analysis after BTK and PI3Kδ inhibitor treatment in chronic lymphocytic leukemia identifies cell-surface CXCR4 as a synergistic target for combinatorial therapy</a></h4>Gabriela Mladonicka Pavlasova, Veronika Sandova, Laura Ondrisova, Vaclav Seda, Petra Pavelkova, Daniel Filip, Krystof Hlavac, Josef Vecera, Sarka Pospisilova, Anna Panovska, Stacey M. Fernandes, Michael Doubek, Jennifer R. Brown, Marek MrazHemaSphere (2026) 10 (9): e70481<a target="_blank" rel="noopener noreferrer" href="https://doi.org/10.1002/hem3.70481">https://doi.org/10.1002/hem3.70481</a><h6>licensed under <a target="_blank" rel="noopener noreferrer" href="https://creativecommons.org/licenses/by/4.0/">CC-BY 4.0</a></h6></blockquote>Marek Mráz’s research group is part of the National Institute for Cancer Research (<a target="_blank" rel="noopener noreferrer" href="https://www.nuvr.cz/en/">NICR</a>).

Leukaemia Cells Respond to Treatment by Turning Themselves into a Target

Are you currently using single-column batch chromatography to isolate and purify target compounds, but struggling with high solvent demand, low product concentration, or limited throughput? Simulated Moving Bed chromatography offers an alternative approach that can address many of these limitations through continuous multi-column separation.<h4>What is Simulated Moving Bed Chromatography (SMB)?</h4>Simulated Moving Bed (SMB) chromatography is a continuous multi-column separation technique designed to make more efficient use of the stationary phase while minimizing solvent consumption. It is particularly well suited to the purification of binary and pseudo-binary mixtures, including enantiomers, sugars, petrochemical products, and fine chemicals.<h4>Conventional Batch Chromatography</h4>In traditional batch chromatography, the sample is introduced onto a single chromatographic column and transported through the stationary phase by the mobile phase. Individual compounds interact with the stationary phase to different extents and therefore travel through the column at different rates:<ul><li>Weakly retained compounds migrate more rapidly.</li><li>Strongly retained compounds move through the column more slowly.</li></ul>The separated substances consequently leave the column at different retention times and can be collected as individual fractions (Fig. 1). Before a new sample is normally introduced, the previous sample must be completely eluted and the column prepared for the next separation.Although single-column batch chromatography is versatile and comparatively easy to operate, several disadvantages become increasingly important at production scale. The stationary phase is not continuously involved in useful separation, which can limit productivity. Other drawbacks include substantial solvent consumption, diluted product fractions, repeated column regeneration, and potential loss of valuable product.<img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/KNAUER_Figure_1_Single_column_separation_A_fast_eluting_component_B_retaining_component_cda8ef66ef.jpg" alt="KNAUER: Figure 1: Single column separation; A – fast eluting component, B – retaining component" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_KNAUER_Figure_1_Single_column_separation_A_fast_eluting_component_B_retaining_component_cda8ef66ef.jpg 76w," sizes="100vw" width="394" height="800"><h4>From True Moving Bed to SMB</h4>The principle behind SMB originates from the theoretical True Moving Bed (TMB) process. In an ideal TMB system, the liquid mobile phase and the solid stationary phase continuously travel in opposite directions. This counter-current arrangement takes advantage of differences in the affinity of individual compounds toward the two phases.Consider a binary mixture containing components A and B. Component A interacts more strongly with the stationary phase, whereas component B is less strongly retained and preferentially follows the liquid mobile phase. Under appropriate operating conditions, the two substances effectively migrate in opposite directions and can be collected continuously through separate outlets (Fig. 2).While this concept is highly efficient from a theoretical perspective, continuously moving a packed solid chromatographic adsorbent is technically difficult to realize.<img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/KNAUER_Figure_2_True_Moving_Bed_concept_e1eac66028.jpg" alt="KNAUER: Figure 2: True Moving Bed concept" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_KNAUER_Figure_2_True_Moving_Bed_concept_e1eac66028.jpg 186w," sizes="100vw" width="954" height="800">Four process ports divide the system into four functional zones (Zone I–IV):<ol><li>Eluent (desorbent) inlet</li><li>Extract outlet</li><li>Feed inlet</li><li>Raffinate outlet</li></ol>Feed and eluent are introduced continuously, while two product streams are simultaneously withdrawn:<ul><li>Extract – the strongly retained, later-eluting component</li><li>Raffinate – the less strongly retained, earlier-eluting component</li></ul><h4>The Four Functional Zones</h4>Each of the four zones has a defined purpose and operates at its own internal flow rate (VI–VIV):<ul><li>Zone I regenerates the stationary phase.</li><li>Zone II prevents the weakly retained component from entering the extract stream.</li><li>Zone III prevents the strongly retained component from contaminating the raffinate.</li><li>Zone IV enables recovery and internal reuse of the remaining solvent.</li></ul><h4>How is the TMB Principle Implemented in SMB?</h4><a target="_blank" rel="noopener noreferrer" href="https://www.knauer.net/smb">SMB</a> eliminates the need to physically move the stationary phase. Instead, the adsorbent remains fixed inside the chromatographic columns, while the positions of the liquid inlet and outlet ports are periodically shifted.From the perspective of these moving connection points, the stationary phase appears to travel in the opposite direction to the liquid phase. SMB therefore reproduces the counter-current behavior of a TMB system without physically transporting the packed adsorbent (Fig. 3).<img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/KNAUER_Figure_3_Transfer_TMB_concept_to_SMB_concept_e4b8905269.jpg" alt="KNAUER: Figure 3: Transfer TMB concept to SMB concept" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_KNAUER_Figure_3_Transfer_TMB_concept_to_SMB_concept_e4b8905269.jpg 245w," sizes="100vw" width="1372" height="800">A typical SMB setup consists of several chromatographic columns arranged in a closed loop. A standard configuration often uses eight columns, although the number can be adapted according to the application.<img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/KNAUER_Figure_4_SMB_concept_fb005cdfa1.jpg" alt="KNAUER: Figure 4: SMB concept" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_KNAUER_Figure_4_SMB_concept_fb005cdfa1.jpg 195w," sizes="100vw" width="1001" height="800">After a predefined switching time, all inlet and outlet positions move forward by one column in the direction of the mobile-phase flow. The columns themselves stay in place, but their functional roles within the process change.By repeatedly shifting the ports, the system creates the effect of a stationary phase moving counter-currently to the mobile phase. With eight columns, eight switching steps are required to complete one full SMB cycle.Following an initial transition period, the process reaches a cyclic steady state. Concentrations continue to fluctuate during individual switching intervals, but the overall concentration profile repeats after every complete cycle.Increasing the number of columns and reducing the size of individual switching steps allows the SMB system to reproduce ideal TMB behavior more closely. Systems containing fewer columns exhibit more pronounced discrete switching effects and can require more sophisticated dynamic modeling during process development.<h3>What are the Main Advantages of SMB?</h3><h4>1. Continuous Operation and Higher Productivity</h4>The key benefit of SMB chromatography is continuous operation. Feed and eluent enter the system continuously, while purified extract and raffinate streams are removed at the same time.As a result, SMB systems can operate for prolonged periods—from several days to even several weeks—with very limited interruption.Because several chromatographic columns perform different separation functions simultaneously, the stationary phase is utilized much more effectively than in conventional batch chromatography. This can translate into higher productivity, improved process efficiency, and a continuous supply of purified product.<h4>2. Reduced Solvent Consumption</h4>Conventional batch chromatography can require substantial volumes of mobile phase to achieve separation and product recovery. The collected fractions are often highly diluted, creating an additional need for downstream concentration.SMB maintains concentration profiles within the circulating system and uses the mobile phase more efficiently. Partial internal recycling of solvent is an inherent part of the process.Lower solvent consumption can lead to:<ul><li>Reduced raw-material costs</li><li>Lower volumes of liquid waste</li><li>Reduced energy requirements for solvent evaporation</li><li>More concentrated product streams and less downstream concentration</li></ul><h4>3. High Recovery and Reproducible Product Quality</h4>Once the process reaches cyclic steady state, SMB can continuously generate product streams with stable and reproducible composition.Additional advantages can include high product recovery, lower product dilution, more efficient use of the stationary phase, and straightforward integration into continuous manufacturing strategies.<h4>What are the Differences Between Batch LC and SMB?</h4><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/KNAUER_differences_between_batch_LC_and_SMB_eba3897f65.jpg" alt="KNAUER: Differences between batch LC and SMB" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_KNAUER_differences_between_batch_LC_and_SMB_eba3897f65.jpg 245w," sizes="100vw" width="1061" height="638"><h4>Where is SMB Used?</h4>SMB technology is established across several industries and can be applied to purification processes ranging from gram-scale production to several tons per day (Fig. 5).Typical and emerging applications include:<ul><li>Sugar purification</li><li>Petrochemical separations</li><li>Chiral separations</li><li>Fine chemical production</li><li>Pharmaceutical intermediates</li><li>Selected biomolecule separations</li></ul><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/KNAUER_Figure_5_SMB_application_areas_7109ff703f.jpg" alt="KNAUER: Figure 5: SMB application areas; author's illustration" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_KNAUER_Figure_5_SMB_application_areas_7109ff703f.jpg 245w," sizes="100vw" width="1500" height="512"><h4>Limitations of SMB</h4>Although SMB can provide substantial advantages, it is not necessarily the ideal solution for every chromatographic separation.The instrumentation and process configuration are more complex than conventional single-column chromatography. Several pumps, valves, columns, and liquid pathways must be controlled precisely. Successful method development therefore requires a solid understanding of adsorption equilibria, mass-transfer effects, and fluid dynamics.Classical four-zone SMB is primarily intended for binary and pseudo-binary separations. More complicated mixtures may require modified SMB configurations, additional purification steps, or multiple coupled SMB systems.SMB is generally most economically attractive when relatively large quantities of a consistent feed material are processed continuously over extended periods.Batch chromatography can remain the better option for small-scale production, frequent changes between products, complex multicomponent mixtures, or applications that depend on flexible gradient separations.<h4>Conclusion</h4>Simulated Moving Bed chromatography converts the theoretical counter-current principle of True Moving Bed separation into a practical continuous chromatographic process. Instead of physically transporting the stationary phase, SMB periodically changes the positions of the feed, eluent, extract, and raffinate connections. This reproduces the effect of counter-current movement between stationary and mobile phases.Compared with conventional batch chromatography, SMB can provide higher productivity, more efficient stationary-phase utilization, substantially lower solvent consumption, and more concentrated product streams. Continuous operation makes the technology particularly attractive for demanding large-scale binary separations, including the purification of pharmaceutical enantiomers.Although successful SMB implementation requires accurate process data, careful method development, and precise control of operating conditions, the technology demonstrates how an idealized chromatographic principle can be transformed into a robust and efficient industrial separation strategy.

Simulated Moving Bed Chromatography: Efficient Continuous Separation

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Leukaemia Cells Respond to Treatment by Turning Themselves into a Target

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Simulated Moving Bed Chromatography: Efficient Continuous Separation