Patented Processes Addresses Pitfalls Associated With Tandem Dye Technology

By Author: Scott eric
Total Articles: 8
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Multicolor flow cytometry analysis is an important tool used in the identification and investigation of cellular subpopulations. Tandem dyes have beendeveloped to expand the portfolio of fluorochromes to meet the growing needs of the research community.

Furthermore, a proprietary and patented process has been developed to provide enhanced utility and quality oftandemfluorochrome dyeproducts and help researches.

Tandem dyes exploit the principle of fluorescence resonance energy transfer (FRET). Since the transfer efficiency depends on the distance between the donor and acceptor chromophore, it is important to conjugate them in close proximity. The patented conjugation process uses a method of intramolecular unfolding of the donor phycobiliprotein (e.g.PE or APC), allowing the acceptor reactive dye molecules to react preferentially with the newly exposed hydrophobic regions containing thebilinchromophores.

Another important aspect of the patented process is the purification of tandem dye molecules by hydrophobic interaction chromatography (HIC). This process has been validated for stringent lot-to-lot reproducibility, ...
... a key requirement for use of antibody conjugates of these dyes in longitudinal or multi-site studies.

Reagents with high background staining of tandem dye acceptor moieties (e.g. Cy5, Cy7, Texas Red) to monocytes pose a major problem when evaluation of dimly-labeled lymphocyte or monocyte populations is critical. However, there is an existing proprietary chemistry in the formulation of products to eliminate this background staining.

Beyond controlling the manufacture of the tandem dyes and the resultant performance of the conjugates at the time of manufacture, it is important to minimize thefluorochromedegradation process that occurs over time with exposure to light.

Bottling in standard amber glass vials that protects the dyes from UV radiation, but not from the visible, and in particular the red, radiation that corresponds to the absorption of both donor and acceptor molecules of most tandem dyes. This has resulted in the development of a unique packaging approach using a black opaque coating to protect tandem dyes from photodegradation. Tandem dye reagents bottled in these black-coated vials and exposed to controlled lighting conditions showed no degradation compared to the amber-vialed product measured by flow cytometry.

In summary, the pitfalls associated with tandem dye technology have been addressed by patented manufacturing processes, formulation chemistry and packaging approaches, assuring delivery of the highest quality dye conjugates which are available in both catalog offerings and custom product portfolio of dye options.

Reference–https://www.beckmancoulter.com/wsrportal/bibliography?docname=Cellular+Research+Issue+1+-+Feb+2007+%28NL-10344A%29.pdf