Proteomic characterization of freeze-dried human plasma: providing treatment of bleeding disorders without the need for a cold chain.
BACKGROUND
Transfusion of human plasma is a basic treatment for severe coagulopathies, especially in major bleeding. The required logistics to provide plasma is challenging because of the need to maintain a cold chain. This disadvantage could be overcome by lyophilized plasma. However, it is unknown to what extent lyophilization alters plasma proteins. Quantitative proteomic technologies were applied to monitor protein changes during production of lyophilized, solvent/detergent (S/D)-treated plasma.


METHODS
The impact of S/D treatment and lyophilization on the plasma proteome was evaluated by differential in-gel electrophoresis (2D-DIGE), and proteins were characterized by mass spectrometry. Clotting factor activities were determined in lyophilized S/D-treated plasma after 24 months of storage at room temperature.


RESULTS
By 2D-DIGE, 600 individual protein spots were compared. Lyophilization did not change any of the 600 spots, whereas pathogen inactivation caused significant changes of 38 spots including alpha1-antitrypsin, alpha1-antichymotrypsin, and alpha2-antiplasmin. Clotting factor activities remained stable over 24 months of storage.


CONCLUSIONS
Lyophilization of human plasma neither alters its protein composition nor impairs its clotting capacity. It does not require cost-intensive logistics for storage and transport and can be quickly reconstituted. It is suggested that lyophilized, pathogen-inactivated plasma is an attractive option to provide the most important basic treatment for severe coagulopathies in areas without cold chain and to provide plasma with reduced time delay in emergency situations.
Steil
Leif
L
Interfaculty Institute for Genetics and Functional Genomics, Institute for Immunology and Transfusion Medicine, Ernst-Moritz-Arndt University, Greifswald, Germany.
Thiele
Thomas
T
Hammer
Elke
E
Bux
Jürgen
J
Kalus
Monika
M
Völker
Uwe
U
Greinacher
Andreas
A
D001793:Blood Preservation / Q000379:methods*; D001797:Blood Protein Electrophoresis; D001798:Blood Proteins / Q000032:analysis* / Q000187:drug effects / Q000627:therapeutic use; D003902:Detergents / Q000494:pharmacology; D015180:Electrophoresis, Gel, Two-Dimensional; D005374:Filtration; D005612:Freeze Drying; D006474:Hemorrhagic Disorders / Q000628:therapy*; D006801:Humans; D013058:Mass Spectrometry; D010949:Plasma / Q000737:chemistry* / Q000187:drug effects / Q000382:microbiology / Q000821:virology; D011489:Protein Denaturation / Q000187:drug effects; D040901:Proteomics; D012034:Refrigeration; D012997:Solvents / Q000494:pharmacology; D013696:Temperature; D013997:Time Factors; D038301:Virus Inactivation
D001798:Blood Proteins; D003902:Detergents; D012997:Solvents