N-methacryloly-(L)-histidinemethylester carrying a pseudospecific affinity sorbent for immunoglobulin-G isolation from human plasma in a column system.
N-methacryloly-(L)-histidinemethylester (MAH) as a pseudospecific ligand was synthesized by using methacryloyl chloride and histidine. Spherical beads with an average size of 63-75 microm were obtained by suspension polymerization of ethylene glycol dimethacrylate (EGDMA), 2-hydroxyethyl methacrylate (HEMA) and MAH conducted in an aqueous dispersion medium. The specific surface area of the beads was found to be 18.3 m2/g. Poly(EGDMA-HEMA-MAH) beads were used in the separation of immunoglobulin-G (HIgG) from aqueous solutions and/or human plasma in a packed-bed column system. HIgG adsorption capacity of the beads decreased with an increase in the flow-rate of plasma. The maximum HIgG adsorption on the poly(EGDMA-HEMA-MAH) sorbents was observed at pH 7.4. HIgG adsorption onto the poly(EGDMA-HEMA) sorbents was negligible. Higher adsorption values (up to 135 mg/g) were obtained when the poly(EGDMA-HEMA-MAH) sorbents were used from aqueous solutions. HIgG adsorption increased with decreasing temperature and the maximum adsorption achieved at 4 degrees C. MAH incorporation significantly affected HIgG adsorption capacity (135 mg/g). Higher amounts of HIgG were adsorbed from human plasma (up to 165 mg/g). Adsorption capacities of other blood proteins were obtained as 8.7 mg/g for fibrinogen and 14.6 mg/g for albumin. The total protein adsorption was determined as 191 mg/g. The pseudospecific affinity beads allowed one-step separation of HIgG from human plasma. HIgG molecules could be repeatedly adsorbed and desorbed with these sorbents without noticeable loss in their HIgG adsorption capacity.
Ozkara
Serpil
S
Bioengineering Division, Hacettepe University, Ankara, Turkey.
Garipcan
Bora
B
Pişkin
Erhan
E
Denizli
Adil
A
D000327:Adsorption; D002846:Chromatography, Affinity / Q000295:instrumentation*; D017743:Equipment Reuse; D006639:Histidine / Q000031:analogs & derivatives / Q000138:chemical synthesis* / Q000737:chemistry / Q000276:immunology; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007074:Immunoglobulin G / Q000097:blood* / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification*; D015394:Molecular Structure; D009994:Osmolar Concentration; D012680:Sensitivity and Specificity; D012996:Solutions; D013438:Sulfhydryl Compounds / Q000138:chemical synthesis* / Q000737:chemistry / Q000276:immunology; D013696:Temperature
D007074:Immunoglobulin G; C486017:N-methacryloly-(L)-histidinemethylester; D012996:Solutions; D013438:Sulfhydryl Compounds; D006639:Histidine