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Although our ferrofluids don’t require a column, individuals wishing to use high gradient techniques (i.e., columns) should be able to do so. Our ferrofluids were designed to obviate the need for either columns or mesh filters, greatly simplifying cell selection procedures.
Using our FerroSelect Quadrupole magnets, selection of cells occurs when the tube is placed inside the magnet. Labeled cells remain attached to the tube wall, and the unlabeled cells can be poured off.
While the reagents offered will be derived from the same clones and starting materials, the cGMP materials will undergo additional testing and full documentation for filing with regulatory agencies. Investigators can undertake development work in their lab and be assured that the same versions of the monoclonal antibodies will be available specifically for clinical use. Wherever possible we use materials that are recombinant and xenofree.
The major benefit of negative depletion (sometimes called negative selection) over positive selection is that recovered cells do not have residual capture reagents on their surface. To achieve cell enrichment through the depletion of unwanted cell types most competitive technologies require a cocktail of antibodies. We accomplish negative depletion with the use of a single mAb that can enrich for CD3 cells and when used in combination with other antibodies, alternative cell subtypes can be enriched.
The cell numbers must be an approximation as patient/donor materials are highly variable. The cell numbers we quote are intended as a guide for investigators to develop their own processes. Our team is available to support you, should you need assistance
The viability of the starting cell population can impact the functionality of any cell separation technology based upon the use of magnetic particles. Poor cell viability introduces the possibility of free DNA being present in the samples. The charged nature of the nucleic acids can cause nonspecific binding. Investigators wishing to process products with low cell viability may wish to include DNase in the selection procedure. Upon request, our team can offer advice regarding what materials may be used
We optimize the number of biotin molecules on our antibodies used in cell selection procedures. Typically, this is in the range of 4-6 biotin molecules per antibody. We can offer investigators protocols for coupling biotin to antibodies, should this be requested. However, we cannot guarantee that these will work under all circumstances. If a biotin molecule attaches within the binding site of the mAb, the antibody will likely be inactivated.
At the end of a selection procedure the “immune capture reagents” are present upon the “positively” selected cells. However, if these cells are placed in culture, after a few days the reagents can no longer be detected using established methods to identify either streptavidin or biotinylated monoclonal antibody. The selection materials either fall off the cells or are internalized.
We have looked at whether the positive selection of T cells using a biotinylated anti-CD3 mAb causes their activation. Using markers that are a surrogate for T cell activation, there is no evidence to suggest that activation occurs in the absence of a separate activation reagent (anti-CD28).