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particles. Incubation is not required, as the gel simply dissolves in less than a minute. Cell vi- ability is significantly increased in comparison to conventional cell separation products, and the cell capture particles that are often retained from isolated cell populations are eliminated. Importantly, the dissolution process does not affect cell phenotype or viability, which makes the isolated cells more suitable for downstream analysis and applications.


Featuring QuickGel technology, the MagCloudz streptavidin cell separation kit provides the high-efficiency capture and label-free release of high-purity target call populations from complex biological samples. The magnetic separation and dissociation process leaves isolated target cells completely label free for scalable downstream culture, analysis and functional studies (see Figure 1).


Achieving complete cell release MagCloudz averages ≥60% recovery of the


initial target cell population and takes advan- tage of the high affinity of streptavidin–biotin interactions to offer biocompatibility, low non- specific binding and high viability of magnetic, label-free, recovered target cells. The process is outlined below:


1. Heterogeneous cell populations are labeled with a biotinylated antibody, directed against the cellular marker of interest (i.e., CD3-biotin for T-cell targeting).


2. Biotin-labeled cell populations are incubated with the appropriate amount of MagCloudz in cell separation buffer; this allows target- cell binding to the MagCloudz surface.


3. Undesired cells are removed by applying a magnetic field to the sample. The target cell–MagCloudz complexes are magnetically separated and the remaining undesired cells are removed in the supernatant.


4. The target cell–MagCloudz complex is rinsed to remove any residual, nonspecifically adhered cells, further increasing target cell purity.


5. The addition of proprietary release buffer gently dissolves the QuickGel, which releases the target cells and removes the magnetic carrier particles from the cells.


6. A magnetic field is again applied to remove the magnetic carrier particles, which leaves the target cells behind in the supernatant.


AMERICAN LABORATORY 41 MARCH 2016


The supernatant containing target cells can then be processed as desired, achieving a highly pure and viable target cell population for further functional studies.


The kits are designed to be used for cells from mammalian sources and can process up to 100 samples containing 10 million (107 each, for a total of 1 billion (109


) cells ) processed


per kit. A maximum of 16 1.5-mL samples can be processed simultaneously using the Q-Mag magnetic stand (Quad Technologies). In some cases, processing times are shorter than conventional techniques, which require approximately two hours.


Case study: CD3+ T-cell isolation The MagCloudz kit has been used success-


fully in several applications, including the isolation of CD3-positive (CD3+) T-cells from peripheral blood fractions. In one study, cell populations were stained with peridinin chlo- rophyll protein complex (PerCP)-conjugated anti-CD3, phycoerythrin (PE)-conjugated anti- CD45, annexin-V-FITC (apoptosis marker) and


propidium iodide (viability marker) following the MagCloudz streptavidin cell separation protocol. Figure 2 shows typical data when using a biotinylated CD-3 antibody from eBio- science (San Diego, Calif.).


Conclusion A cell separation solution that delivers highly pure and viable target cells that are magnetic label-free will significantly benefit critical research applications, drug development and cancer treatments such as stem-cell transplants and immune therapy. Researchers can easily scale up their workflows to increase through- put without compromising yield, efficiency, viability or purity.


Andrea Armstead, Ph.D., is technical market- ing specialist, Quad Technologies, 3F Gill St., Woburn, Mass. 01801, U.S.A.; tel.: 339-927-9663; e-mail:aarmstead@quadtechnologies.com; www. quadtechnologies.com


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