Evaluating the Potential of Kode Technology to Attach Nucleic Acids onto Red Cells and Create Reticulocyte Mimics
Reticulocytes are immature red cells that still have nuclei acid in them. They are an essential clinical indicator as an increase in their number indicates rapid turn-over of red cells. Most advanced automated haematology machines include a diagnostic assay for reticulocytes, and all diagnostic assays require quality control procedures. Herein lies the issue, sourcing cells suitable as controls for reticulocyte QC is problematic. Kode technology uses chemical constructs to modify cell surfaces with known quantities of small molecules. The opportunity exists to establish if human red blood cells can be labelled with Kode constructs that secondarily capture nucleic acids and create surrogate reticulocytes suitable for quality control. This study aimed to investigate the potential to create reticulocyte controls prepared with Kode technology as an alternative to natural reticulocyte controls. Established methods were used to prepare biotin kodecytes and biotinylated DNA and these were attached to biotin-kodecytes via streptavidin. Due to limitations in materials, surrogates for biotinylated DNA were also used to evaluate steps designed to prepare mimic reticulocytes. A haematology analyser and supravital staining were used to assess the kodecyte mimic reticulocytes. This research had partial success. DNA was successfully extracted, fragmented and biotinylated. Methods demonstrating that biotinylated polymers, including biotinylated DNA, could be attached to red cells koded with biotin via streptavidin bridge was also successful. However, the final product of mimic reticulocytes (biotin-kodecytes + streptavidin + biotinylated DNA fragments) was not successful.
Kode Technology, Reticulocyte, Biotinylated DNA, Streptavidin
Networked Digital Library of Theses and Dissertations