Cell Surface Modification

FSL Constructs for Labeling of Cells and Real-Time Imaging

Rapid Nontoxic in vitro/in vivo Labeling Without Loss of Function or Vitality

FSL Constructs are compounds based on KODE technology designed to label hydrophobic surfaces including living cells with carbohydrates, biotin, fluorophores, or 125I.1 All KODE FSL Constructs consist of three essential designable features:
  • a functional component (F)
  • a spacer (S)
  • and a diacyl lipid (L)
FSL Constructs disperse in water for biocompatibility and spontaneously and stably incorporate into cell membranes.
Cells modified with KODE constructs are known as kodecytes2 and kodevirions3 and are still vital and functional after labeling. Kodecytes and kodevirions retain their normal functionality while gaining the functionality introduced by the FSL Construct. FSL Constructs can also be used to modify other hydrophobic surfaces including fixed cells and solid-phase surfaces including silica, nitrocellulose, and paper.4

FSL Constructs have been used in vitro to modify embryos, spermatozoa, zebrafish5, epithelial/endometrial cells, red blood cells,2,6 and virions.3,7 Kodecytes have been infused intravascularly into experimental animals in vivo without loss of vitality.8,9

An improved method to screen Fc function of immunoglobulin products was developed using CMV kodecytes,10 and FSL antigen constructs have been printed onto silica to create a convenient array for antibody identification.4 Measles virions (MV) were fluorescently labeled to monitor binding to DC-SIGN+ expressing CHO cells using flow cytometry.7 Carbohydrate FSL Constructs11 have been used to modify cells and viruses to study virus inhibition,12 identify Lewis antibody,13 prepare labeled cells for ABO QC methods,2,13 and tag cells for flow cytometry.6
The combination of dispersibility in biocompatible media, spontaneous incorporation into cell membranes, and apparent low toxicity, makes FSL Constructs a valuable research tool for the development of new cell biology applications.

View the Journal of Visualized Experiments (JoVE) video on (KODE technology).
References
1. FSL Constructs: A simple method for modifying cell/virion surfaces with a range of biological markers without affecting their viability. Blake, D.A., Bovin, N.V., Bess, D., and Henry, S.M., J. Visualized Experiment (JoVE), 54, e3289 (2011) (Video article).
2. Henry, S.M., Modification of red blood cells for laboratory quality control use. Curr. Opin. Hematol. 16, 467-72 (2009).
3. Fluorescein and radiolabeled Function-Spacer-Lipid constructs allow for simple in vitro and in vivo bioimaging of enveloped virions. Hadac, E.M., Federspiel, M.J., Chernyy, E., Tuzikov, A., Korchagina, E., Bovin, N.V., Russell, S. and Henry, S.M., J. Viriological Methods, 176, 78-84 (2011).
4. Inkjet printing of Function-Spacer-Lipid (FSL) biomolecules, Barr, K., Diegel, O., Parker, S., and Henry, S., NZ Rapid Product Development Conference 2011, Auckland University of Technology, Auckland, New Zealand.
5. Fluorescent Function-Spacer-Lipid construct labeling allows for real-time in vivo imaging of cell migration and behavior in zebrafish. Lan, C.-C., Blake, D., Henry, S., and Love, D.F., J. Fluorescence, 22, 1055-63 (2012).
6. Flow cytometry evaluation of red blood cells mimicking naturally-occurring ABO subgroups following modification with variable amounts of FSL-A and B constructs. Hult, A.K., Frame, T., Chesla, S., Henry, S., and Olsson, M.L. Transfusion, 52, 247-51 (2011).
7. A prominent role for DC-SIGN+ dendritic cells in initiation and dissemination of measles virus infection in non-human primates. Mesman, A.W., de Vries, R.D., Duprex, W.P., de Swart, R.L., and Geijtenbeek, T.B.H., PLoS One, 7, e49573 (2012).
8. Modeling transfusion reactions and predicting in vivo cell survival with kodecytes. Oliver, C., Blake, D., and Henry, S., Transfusion, 51, 1723-30 (2011).
9. In vivo neutralization of anti-A and successful transfusion of A antigen-incompatible red blood cells in an animal model. Oliver, C., Blake, D., and Henry, S., Transfusion, 51, 2664-75 (2011).
10. An improved Fc function assay utilizing CMV antigen-coated red blood cells generated with synthetic function-spacer-lipid constructs. Georgakopoulos, T., Komarraju, S., Henry, S., and Bertolini, J., Vox Sanguinis, 102, 72-8 (2012).
11. Toward creating cell membrane glyco-landscapes with glycan lipid constructs. Korchagina, E., Tuzikov, A., Formanovsky, A., Popova, I., Henry, S., and Bovin, N., Carbohydrate Res., 356, 238-46 (2012).
12. A synthetic globotriaosylceramide analogue inhibits HIV-1 infection in vitro by two mechanisms. Harrison, A.L., Olsson, M.L., Brad Jones, R., Ramkumar, S., Sakac, D., Binnington, B., Henry, S., Lingwood, C.A., and Branch, D.R., Glycoconjugate J., 27, 515-524 (2010).
Image of murine embryo labeled with FSL-biotin (Catalog No. F9182) and detected using fluorescent
Image of 8 day zebrafish after sinus venosus infusion using FSL-fluorescein (Catalog No. F1058) to

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F9182 FSL-biotin All FSL constructs disperse in biocompatible media and spontaneously and stably incorporate into cell membranes. Cells modified with FSL constructs are known as kodecytes and usually maintain their normal vitality and functionality. FSL-biotin has been specifically designed to create biotinylated live cells (kodecytes) but can also be used to modify other hydrophobic surfaces including fixed cells, virions, and solid phase surfaces. Biotinylated kodecytes can be reacted with a variety of biotin binding protein constructs including fluorescent labeled avidin and avidinylated beads/solid surfaces.
F1058 FSL-Fluorescein All FSL constructs disperse in biocompatible media and spontaneously and stably incorporate into cell membranes. Cells modified with FSL constructs are known as kodecytes and usually maintain their normal vitality and functionality. FSL-fluorescein has been specifically designed to create fluorescein-labeled live cells (kodecytes) but can also be used to modify other hydrophobic surfaces including fixed cells, virions, and solid phase surfaces. Fluorescein-labeled kodecytes can be visualized at the approximate absorbance wavelength of 488 nm and emission at 518 nm.
F9432 FSL-Galili(tri) All FSL constructs disperse in biocompatible media and spontaneously and stably incorporate into cell membranes. Cells modified with KODE constructs are known as kodecytes and usually maintain their normal vitality and functionality.

FSL-Galili(tri) has been specifically designed to create human cells (kodecytes) expressing controlled levels of the animal Galili antigen but can also be used to modify other hydrophobic surfaces including fixed cells, virions, and solid phase surfaces. This product can be used to modify Galili negative red cells to create kodecytes that can quantitate antibodies directed against the Galili antigen. These Galili-kodecytes will react with almost all human serum, the degree of reactivity determined by the level of antibody present (and level of FSL antigen inserted).
F9807 FSL-GB3 All FSL constructs disperse in biocompatible media and spontaneously and stably incorporate into cell membranes. Cells modified with KODE constructs are known as kodecytes and usually maintain their normal vitality and functionality.

FSL-GB3 has been specifically designed to create live cells (kodecytes) expressing GB3 antigens but can also be used to modify other hydrophobic surfaces including fixed cells, virions, and solid phase surfaces. This product has been used in vitro to inhibit HIV virions and bind to veratoxin.
F9682 FSL-LEa(tri) All FSL constructs disperse in biocompatible media and spontaneously and stably incorporate into cell membranes. Cells modified with KODE constructs are known as kodecytes and usually maintain their normal vitality and functionality.

FSL-Lea(tri) has been specifically designed to create live cells (kodecytes) expressing blood group Lea antigens but can also be used to modify other hydrophobic surfaces including fixed cells, virions, and solid phase surfaces. This product can be used to attach reproducible and controlled levels of blood group Lea antigens to erythrocytes. These kodecytes will react with most anti-Lea blood grouping reagents including polyclonal reagents. Lea-kodecytes will also react with monoclonal anti-Leb as these reagents also cross-react with Lea.
F9307 FSL-A(tri) All FSL constructs disperse in biocompatible media and spontaneously and stably incorporate into cell membranes. Cells modified with KODE constructs are known as kodecytes and usually maintain their normal vitality and functionality.

FSL-A(tri) has been specifically designed to create live cells (kodecytes) expressing blood group A antigens but can also be used to modify other hydrophobic surfaces including fixed cells, virions, and solid phase surfaces. This product can be used to attach reproducible and controlled levels of blood group A antigens (generic A due to its trisaccharide structure) to group O or B erythrocytes. These kodecytes will react with most anti-A blood grouping reagents including polyclonal reagents
F9557 FSL-B(tri) All FSL constructs disperse in biocompatible media and spontaneously and stably incorporate into cell membranes. Cells modified with KODE constructs are known as kodecytes and usually maintain their normal vitality and functionality.

FSL-B(tri) has been specifically designed to create live cells (kodecytes) expressing blood group B antigens but can also be used to modify other hydrophobic surfaces including fixed cells, virions, and solid phase surfaces. This product can be used to attach reproducible and controlled levels of blood group B antigens (generic B due to its trisaccharide structure) to group O or A erythrocytes. These kodecytes will react with most anti-B blood grouping reagents including polyclonal reagents.
F7558 FSL-Tyr1 ≥90% (TLC) All FSL constructs disperse in biocompatible media and spontaneously and stably incorporate into cell membranes. Cells and virions modified with FSL constructs are known as kodecytes and kodevirions, respectively, and usually maintain their normal vitality and functionality. FSL-Tyr1 has been specifically designed to create radiolabeled (125I) live cells (kodecytes) but can also be used to modify other hydrophobic surfaces including fixed cells, virions, and solid phase surfaces.