Simultaneous depletion of immunoglobulin G and albumin from human plasma using novel monolithic cryogel columns.
In this study, we aimed to develop an alternative matrix able to deplete the albumin (Alb) and immunoglobulin G (IgG) from blood plasma simultaneously to prepare plasma samples for large-scale applications of blood-related proteomics. As a first step, nano-protein A nanoparticles (nanoProA) were prepared and characterized. Subsequently, cibacron blue F3GA (CB) was immobilized onto the nanoProA's to enhance their specific affinity for Alb molecules. Finally, both nanoparticles, specifically, nanoProA and CB-nanoProA, were separately embedded into cryogel structures to combine advantages of the nanoparticles with those of the cryogels. The protein adsorption was optimized using aqueous Alb and IgG solutions separately. Subsequently, competitive protein adsorption was performed using a protein mixture prepared with Alb and IgG adhering to their plasma protein ratios. Because of the CB-immobilization, the Alb depletion performance of the cryogels increased whereas the IgG depleting performance decreased. Using the nanoProA, embedded cryogel removed 99.3% of the IgG, while using the CB-nanoProA embedded cryogel removed 97.5% of the Alb content. The simultaneous depletion performances of the cryogels for Alb and IgG were characterized using sodium dodecyl sulfate-polyacrylamide gel electrophoresis. In this study, the monolithic cryogel-based adsorbents were classified as an alternative matrix to prepare plasma samples for proteomics applications at the preparative scale.
Hacettepe University, Department of Chemistry, Beytepe, Ankara, Turkey. Electronic address: lokman@hacettepe.edu.tr.
Lokman
L
Uzun
Canan
C
Armutcu
Özlem
Ö
Biçen
Arzu
A
Ersöz
Rıdvan
R
Say
Adil
A
Denizli
D000327:Adsorption; D059985:Cryogels / Q000737:chemistry; D006801:Humans; D055767:Immobilized Proteins; D007074:Immunoglobulin G / Q000097:blood* / Q000302:isolation & purification*; D053758:Nanoparticles / Q000737:chemistry / Q000648:ultrastructure; D011102:Polyhydroxyethyl Methacrylate; D040901:Proteomics / Q000379:methods; D012709:Serum Albumin / Q000302:isolation & purification*; D013205:Staphylococcal Protein A; D014227:Triazines
D059985:Cryogels; D055767:Immobilized Proteins; D007074:Immunoglobulin G; D012709:Serum Albumin; D013205:Staphylococcal Protein A; D014227:Triazines; D011102:Polyhydroxyethyl Methacrylate; C012842:Cibacron Blue F 3GA
Albumin; Cibacron blue F3GA; Cryogel; IgG; Protein A; Simultaneous depletion