Abstract
This conceptual framework discloses, as defensive prior art (CC BY 4.0), a near-critical supercritical-CO2 (scCO2) desiccation protocol for ambient-temperature preservation of genetically engineered triple-knockout (TKO) porcine red blood cells (pRBCs), in which the GGTA1, beta4GalNT2, and CMAH carbohydrate-antigen genes are inactivated and human complement-regulatory transgenes (e.g. CD47, CD55) are expressed. No experimental haemolysis, recovery, or potency data are generated; all values are cited literature anchors. TKO pRBCs are described accurately as genetically advanced but preclinical (Yamamoto 2021; Ko 2022; Chornenkyy 2023), with no clinically deployed pig-red-cell product and current handling assuming a cold chain. The protocol operates at 74-80 bar and 31-35 C, just above the CO2 critical point (31.1 C, 73.8 bar), where near-zero surface tension permits drying with, by hypothesis, minimised membrane perturbation. The central tension is stated explicitly: scCO2 is also a documented agent of microbial inactivation and membrane disruption (Tamburini 2014), so its use here as a gentle drying medium is a near-critical-bounded hypothesis, not a demonstrated capability. The principal barrier remains post-rehydration haemolysis: the best published desiccation results are roughly an order of magnitude above the 0.8% limit (Council of Europe/EDQM). Candidate mitigations are analysed (intracellular trehalose vitrification; controlled rehydration osmolarity; complement-regulator retention). Validation (XRPD/DSC, 24-hour recovery, haemolysis
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Recommended Citation
chuenglertsiri, chaiyapat and Chen, Pattrick, "Near-Critical Supercritical-CO₂ Desiccation of Triple-Knockout Porcine Red Blood Cells: A Proposed Protocol with Haemolysis-Reduction Pathway Analysis", Technical Disclosure Commons, ()
https://www.tdcommons.org/dpubs_series/10485