The CliniMACS System - Immunomagnetic Cell Separation in a Closed, Sterile Environment

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Miltenyi Biotec, founded in 1989, has become one of the most successful biotechnology companies in Germany, and is a premier provider of products and services for integrated cell research. Miltenyi Biotec has established the renowned MACS® Technology as the gold standard in magnetic approaches to the separation of cells - the ease and speed of which comprise only two of its many advantages. The CliniMACS Cell Selection System, based upon MACS Technology, takes magnetic cell separation one step further, providing a closed, sterile system for the enrichment of specific cell populations that can subsequently be used in clinical research applications.

All higher organisms are composed of many different cell types with a plethora of different functions. For research and biomedical applications in immunology, haematology, cell biology and molecular biology, there is often the need to isolate a specific population of cells. With MACS Technology, Miltenyi Biotec offers and develops powerful products that enable the specific enrichment of a variety of cell populations. MACS Technology can be used for the isolation of virtually any cell type from humans or animals.

Principle of MACS Technology

MACS Technology is based on the use of MACS MicroBeads - nano-sized superparamagnetic particles coupled to specific antibodies - patented MACS Columns, and MACS Separators-strong permanent magnets. MACS MicroBeads are approximately 50nm in diameter and are composed of an iron dextran matrix, which has an excellent safety profile. In our experience, MACS MicroBeads do not alter the structure, function or activation status of labelled cells and are not known to interfere with subsequent experiments.

As depicted in Figure 1, cell separation with MACS Technology takes place within MACS Columns when placed in a MACS Separator. Magnetically labelled cells are retained within the column. The MACS Column matrix provides a magnetic field strong enough to retain target cells that are labelled with even minimal amounts of MACS MicroBeads. Unlabelled cells pass through and can be collected; labelled cells are released and collected after removal of the magnetic field.

The CliniMACS System

Using the principles of MACS Technology, the CliniMACS Plus Instrument (Figure 2) has been developed to permit the automated separation of cells on a clinical-scale level in a closed and sterile system. Magnetic selection or depletion of labelled target cells, for example from leukapheresis product, can be performed to isolate a desired cell population, or for the specific removal of a given cell population, based on the recognition of a surface antigen by highly specific antibodies.

The key components of the CliniMACS Plus Instrument, as outlined in Figure 3, are the integrated microcomputer, the magnetic separation unit, the peristaltic pump and various pinch valves. The integrated computer controls all components of the instrument and directs the system to perform repeated procedures in a standardised sequence. The magnetic separation unit includes a movable permanent magnet and a holder for the selection column. The peristaltic pump controls the flow rate throughout the tubing set and, together with the pinch valves, ensures the controlled flow of buffer through the system and continual suspension of cells.

For CliniMACS Cell Separation, target cells are magnetically labelled in the Cell Preparation Bag (see Figure 2) using the CliniMACS Reagent. Like MACS MicroBeads, the CliniMACS Reagent consists of antibody-coupled iron dextran nanoparticles but is supplied in a sterile, non-pyrogenic solution. After labelling, the cells are washed to remove excess reagent. The Cell Preparation Bag is then connected to the Tubing Set, which in turn is connected to the Buffer Bag and the Cell Collection Bag. Tubing Sets consist of pre-assembled sterile tubing, including a pre-column and a separation column, and are for single use only. After initiation of the separation programme, the system automatically applies the cell sample onto the separation column in the field of a CliniMACS Magnetic Separation Unit. Labelled cells are retained within the column, while unlabelled cells are removed by a series of washing steps. Target cells are eluted from the column after removal of the magnetic field, and are collected in high purity within the Cell Collection Bag.

The CliniMACS Plus Instrument, in combination with the CliniMACS Tubing Sets, CliniMACS Reagents and CliniMACS PBS/EDTA Buffer, can separate large numbers of cells and provide target cells within high purity and with excellent yield. It also offers a flexible platform to enable the separation of many cell types using customised separation protocols according to specific sample requirements. Unique Reagents and Tubing Sets also allow for the transition of promising research applications to innovative cell therapy approaches in clinical research.

Clinical Research Applications

The components of the CliniMACS System are CE-marked for clinical use in Europe, and the system is currently being used for the separation of a variety of human cell types. In the US, CliniMACS products are available only under an approved Investigational New Drug (IND) application or Investigational Device Exemption (IDE) for clinical research use. The portfolio of CliniMACS Reagents includes those for the separation of stem and progenitor cells, monocytes, natural killer (NK) cells, B-cells, and T-cell subsets. The CliniMACS Anti-Biotin Reagent also permits a flexible labelling system where biotinylated antibodies can be used to target other populations of cells in a user-defined fashion.

The CliniMACS System has thus far been successfully used in a variety of clinically oriented research applications, mostly within the field of haematology. However, other areas have recently been explored, such as the enrichment of CD133+ stem and progenitor cells for clinical research applications in cardiovascular medicine. The ability of the CliniMACS System to isolate cells directly from different sources, such as leukapheresis product or bone marrow aspirate, facilitates the rapid enrichment of a variety of haematopoietic cell types, including stem and progenitor cells and antigen-specific T-cells.

Together with experienced researchers and clinicians, Miltenyi Biotec is continually developing and refining tools for application within both basic and clinical research, expanding technological boundaries in the field of cell separation and analysis, and responding to the demands of researchers dedicated to cellular research and therapies.

The CliniMACS System components (Reagents, Tubing Sets, Instruments and PBS/EDTA Buffer) are manufactured and controlled under an International Organisation for Standardisation (ISO) 13485 certified quality system. In Europe, the CliniMACS System components are available as CE-marked devices.

In the US, the CliniMACS System components, including the CliniMACS Reagents, are available for use only under an approved Investigational New Drug (IND) application or Investigational Device Exemption (IDE). CliniMACS MicroBeads are for research use only and not for use in humans.

CliniMACS is a registered trademark of Miltenyi Biotec GmbH.Ôûá