Protein depletion from blood plasma using a volatile buffer.
Removal of high abundance proteins is widely used in sample processing for proteomics studies of blood plasma. Immunoaffinity (IA) depletion is currently the most specific method for performing this step. Historically, IA depletion matrices have been designed to be used with inorganic buffers. However, the presence of salts in depleted samples presents a particular problem, and these must be removed in order to make samples compatible with post-depletion processing. Desalting (dialysis, ultrafiltration, size-exclusion, etc.) usually diminishes sample integrity due to labware associated losses. Moreover, these steps require additional labor, increasing the processing time and cost of analysis. In order to avoid these problems, we have developed an IA method using a volatile buffer that can be removed from depleted samples by lyophilization. This method allows the execution of reproducible and efficient depletion of blood plasma in a semi-automated manner.
Sitnikov
Dmitri
D
Caprion Pharmaceuticals, Inc., Department of Protein Analysis, 7150 Alexander Fleming, Montreal, Que., Canada H4S-2C8. dsitnikov@caprion.com
Chan
Donovan
D
Thibaudeau
Eric
E
Pinard
Marc
M
Hunter
Joanna M
JM
D001798:Blood Proteins / Q000302:isolation & purification*; D002021:Buffers; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D005612:Freeze Drying; D006801:Humans; D010949:Plasma; D015203:Reproducibility of Results; D014462:Ultrafiltration
D001798:Blood Proteins; D002021:Buffers