Modeling transfusion reactions with kodecytes and enabling ABO‐incompatible transfusion with function‐spacer‐lipid constructs

Abstract

Background KODE technology uses Function‐Spacer‐Lipid (FSL) constructs for artificial attachment of blood group antigens onto live cells (kodecytes) and visualisation, thus making it potentially suitable to model both transfusion reactions and determine in vivo cell survival. Additionally, as some FSL constructs are glycolipid‐like, the possibility exists that they may also neutralise circulating antibodies, in the same way Lewis glycolipids were historically used to allow Lewis incompatible transfusions. Aims To determine if kodecytes can be used to both mimic and monitor transfusion reactions in an animal model, and also to determine if FSL construct infusions can neutralise circulating antibodies and allow for an incompatible transfusion. Methods Naïve, anti‐A positive (immunised), and anti‐A positive/FSL‐A neutralised mice were given transfusions of murine kodecytes created with FSL‐biotin and FSL‐A. Levels of surviving kodecytes were determined at multiple time‐points via their biotin label. Analysis for anti‐A was performed with inkjet printed FSL‐A constructs in a micro immunoassay. Results Normal kodecyte survival was observed in control animals. Mice with anti‐A predominantly cleared their incompatible A‐kodecyte transfusions within 6 min. Mice with anti‐A that had been neutralised with an FSL‐A infusion had normal survival of their incompatible A‐kodecyte transfusions. Re‐challenge transfusions of A‐kodecytes into mice previously given FSL‐A (once the FSL‐A had cleared from their circulation) gave two different results. One group cleared the incompatible A‐kodecytes within minutes while the other had normal survival, suggesting potential induction of tolerance. Conclusions Kodecytes can be used to model blood group A incompatible transfusions in animals. FSL‐A can neutralise anti‐A and allow for incompatible A‐kodecyte transfusion. The potential exists to extend these research observations into clinical use to allow for transfusion and transplantation across the ABO barrier, or at least mitigate the consequences of accidental ABO incompatibilities.

Ultra-Fast Glyco-Coating of Non-Biological Surfaces

“…However the interaction is not robust, and captured red cells can be released from the microspheres by subjecting them to shear forces such as agitation (e.g., vortex) or aggressive washing procedures. A range of different microspheres including paramagnetic beads [ 10 ] have been modified with FSL constructs.…”

“…Each surface or prepared microplate well was flooded or filled with IgM monoclonal anti-A (Epiclone anti-A, Cat# 0261, CSL, Seaford, VIC, Australia) diluted 1:4 with BSA-PBS and then incubated for 30 min at RT. This anti-A reagent has been previously established to show specific binding against the blood group A antigen, including in the form of FSL constructs [ 4 , 5 , 7 , 8 , 10 , 11 ]. Unprinted areas within the assay are internal blanks.…”

National Blood Foundation 2021 Research and Development summit: Discovery, innovation, and challenges in advancing blood and biotherapies

“…Such modification to the erythrocyte has been done by Kode technology using F unctional head, S pacer, and L ipid (FSL) constructs 22–25 . The lipid is necessary for integration into the cell membrane while the different spacers facilitate a variety of conformational and functional properties.…”

“…Such modification to the erythrocyte has been done by Kode technology using Functional head, Spacer, and Lipid (FSL) constructs. [22][23][24][25] The lipid is necessary for integration into the cell membrane while the different spacers facilitate a variety of conformational and functional properties. The functional head of peptide epitopes is typically less than 20 amino acids and specifically selected to represent the most antigenic regions of a protein; which improves both sensitivity and specificity.…”