Separation of B and T lymphocytes by cellular adsorption chromatography with polyamine graft copolymers as column matrices. II. Recovery of adsorbed B cell enriched populations from the column.
Poly(2-hydroxyethyl methacrylate)-graft-polyamine copolymers were developed for column adsorbents for separating lymphocyte subpopulations collected from rat mesenteric lymph nodes. Bead-shaped adsorbents were prepared by coating poly(2-hydroxyethyl methacrylate)-graft-polyamine copolymer on glass beads. Separation features of the column packed with poly(2-hydroxyethyl methacrylate)-graft-polyamine copolymer-coated glass beads were evaluated under various operating conditions. Separation efficacy, AB/AT, was significantly affected by the infusion rate of lymphocyte suspension into the column, increasing with decreasing infusion rate. T cell purity in the column effluent was almost 95% at the infusion rate of 0.1 ml/min. The infusion rate was also found to affect the detachment of adsorbed lymphocytes from the poly(2-hydroxyethyl methacrylate)-graft-polyamine copolymer surface by mechanical pipetting; lymphocytes adsorbed at a low infusion rate were able to be detached quantitatively by suspending the lymphocyte-adsorbed beads with gentle pipetting. The detached lymphocytes thus obtained were confirmed to be enriched in B cells 1.4 times the initial mixture of B and T cells. The addition of albumin in the medium was found to affect the processes of adsorption and recovery of lymphocyte.
Maruyama
A
A
Department of Industrial Chemistry, Faculty of Engineering, Science University of Tokyo, Japan.
Tsuruta
T
T
Kataoka
K
K
Sakurai
Y
Y
D000327:Adsorption; D000818:Animals; D001402:B-Lymphocytes / Q000187:drug effects*; D002449:Cell Aggregation / Q000187:drug effects; D002469:Cell Separation; D002846:Chromatography, Affinity; D008863:Microspheres; D011073:Polyamines / Q000494:pharmacology*; D011102:Polyhydroxyethyl Methacrylate / Q000494:pharmacology* / Q000031:analogs & derivatives; D011109:Polymethacrylic Acids / Q000494:pharmacology*; D051381:Rats; D011919:Rats, Inbred Strains; D013601:T-Lymphocytes / Q000187:drug effects
D011073:Polyamines; D011109:Polymethacrylic Acids; C056971:poly(2-hydroxyethyl methacrylate)-polyamine graft copolymer; D011102:Polyhydroxyethyl Methacrylate