Increasing storage stability of freeze-dried plasma using trehalose

Abstract

Preservation of blood plasma in the dried state would facilitate long-term storage and transport at ambient temperatures, without the need of to use liquid nitrogen tanks or freezers. The aim of this study was to investigate the feasibility of dry preservation of human plasma, using sugars as lyoprotectants, and evaluate macromolecular stability of plasma components during storage. Blood plasma from healthy donors was freeze dried using 0−10% glucose, sucrose, or trehalose, and stored at various temperatures. Differential scanning calorimetry was used to measure the glass transition temperatures of freeze-dried samples. Protein aggregation, the overall protein secondary structure, and oxidative damage were studied under different storage conditions. Differential scanning calorimetry measurements showed that plasma freeze-dried with glucose, sucrose and trehalose have glass transition temperatures of respectively 72±3.4˚C, 46±11˚C, 15±2.4˚C. It was found that sugars diminish freeze-drying induced protein aggregation in a dose-dependent manner, and that a 10% (w/v) sugar concentration almost entirely prevents protein aggregation. Protein aggregation after rehydration coincided with relatively high contents of β-sheet structures in the dried state. Trehalose reduced the rate of protein aggregation during storage at elevated temperatures, and plasma that is freeze-dried plasma with trehalose showed a reduced accumulation of reactive oxygen species and protein oxidation products during storage. In conclusion, freeze-drying plasma with trehalose provides an attractive alternative to traditional cryogenic preservation.

Ex vivo hemostatic and immuno‐inflammatory profiles of freeze‐dried plasma

“…81 The current findings demonstrating that the relatively low concentrations of immuno-inflammatory proteins present in FP were preserved in FPD, are consistent with the literature suggesting that freeze-drying or lyophilizing processes do not greatly enhance or disrupt the macromolecular stability of commonly detectable inflammatory cytokines, chemokines and other bioactive mediators. 21,84,85 Similarly, results from experimental animal models, administering dried or lyophilized plasma, indicate that FDP likely does not exacerbate existing harmful immune-inflammatory dysregulation after trauma and may in fact, confer beneficial anti-inflammatory effects that result in less blood loss and subsequent organ dysfunction. 25,[86][87][88] Moreover, findings from pre-clinical models of hemorrhage and resuscitation do not indicate that in vivo or ex vivo use of FDP adversely affects the functional interplay between coagulation and inflammatory systems.…”

Freeze-Drying of Platelet-Rich Plasma: The Quest for Standardization

“…In this context, platelets can be preserved by introducing trehalose, a commonly used lyoprotectant, into their cytosol, and freeze-drying in trehalose/albumin buffer [10]. Trehalose is also efficient as an extracellular lyoprotectant, preventing protein aggregation and enhancing plasma stability during storage, and is preferred over sucrose and glucose [11].…”

Room-temperature Storage of Lyophilized Engineered Bacteria using Tardigrade Intrinsically Disordered Proteins

“…Trehalose is commonly used to protect diverse organisms and even enzymes [6,10,15,28,30,34,35] under conditions of desiccation [7,26] . However, it is not present in tardigrades, which are some of the most desiccation-resistant organisms on the planet [8,14,22,33] .…”

“…Secretory-abundant heat soluble protein 33020 from H. dujardini [5,44] LEA Late embryogenesis abundant protein [27,31,46] OtsA trehalose-6-phosphate synthase [6,34,42,47] OtsB trehalose phosphatase [6,34,42,47] Supplementary Figure 1: TDPs were expressed in E. coli BL21 (DE3) using a modified pET28a+ vector.…”