Magnetic bead sorting is an experimental technology based on the specific binding of magnetic particles (magnetic beads) to the target substance, and the target substance is quickly separated and purified by an external magnetic field. This technology is widely used in the fields of biology, chemical analysis, medical diagnosis, etc., especially in the isolation and purification of nucleic acids, proteins, cells, etc.
1. Basic Principle Magnetic bead sorting uses the specific binding between magnetic beads (magnetic beads) and target substances (such as specific antigens on the cell surface) to achieve the separation and purification of target substances under the action of an external magnetic field. Magnetic beads are usually made of iron oxides (such as Fe3O4) or other magnetic materials, with specific isolated substances (such as antibodies, affinity ligands, etc.) on the surface to increase their affinity with the target substance.
2. Operation steps The steps of magnetic bead sorting generally include sample processing, magnetic bead binding, separation and elution: 1. Sample treatment: Pretreat the cell sample, such as cell lysis, protein extraction, etc., to release the target substance or put the cells in a suitable sorting state. 2. Magnetic bead binding: Add magnetic beads modified with specific antibodies to the sample. After thorough mixing, the magnetic beads specifically bind to the target substance (such as specific antigens on the cell surface). 3. Separation: Through the application of an external magnetic field, the magnetic beads and their combined target substances are concentrated to a certain position, thereby achieving the separation of the target substance and the non-target substances. 4. Elution: When necessary, the bond between the magnetic beads and the target substance is destroyed by changing conditions (such as pH, ion concentration, etc.), thereby eluting the pure target substance.
3. Sorting method. Magnetic bead sorting can be performed using a tube-based or column-based method: 1. Tube-based magnetic separation method: Place the suspension of magnetically labeled cells in a magnet, allowing the labeled cells to move near the magnet and be adsorbed, and then perform separation. 2. Magnetic separation method based on column: Separate the suspension of magnetically labeled cells through the separation column in the magnet. The magnetic field in the separation column captures the labeled cells, while the unlabeled cells flow out with the liquid.
4. Sorting type According to the content that is retained or isolated, magnetic bead sorting can be divided into two ways: positive sorting and negative sorting: 1. Positive sorting: Select the cells that need to be collected as the target population, use antibodies to directly label the target cells, and use external magnetic field to make these cells adsorb to the separation column or test tube wall, thereby achieving separation. This method is fast, simple to operate, high purity and high recovery rate, and is suitable for the case where the target cells have surface markers with strong specificity. 2. Negative sorting: Indirectly obtain target cells by labeling non-target cells and removing them. This approach avoids cell activation problems that may arise from direct labeling of target cells and is suitable for situations where specific cell subpopulations or the absence of specific antibodies on the surface of target cells is required.
5. Application fields Magnetic bead sorting technology is widely used in medicine, biology, drug research and development and other fields, such as isolating and purifying biological molecules such as DNA, RNA, protein, and performing immunoassays such as enzyme-linked immunosorbent assays (ELISA), and isolating specific cell types. In cell biology and clinical studies, magnetic bead sorting has become a standard method for high-quality cell sorting.
Magnetic bead sorting is an efficient, specific and widely used cell separation technology. Its principles and operating steps are relatively simple and clear. Different sorting methods and types can be selected according to specific needs.
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