title: Isolation of human serum immunoglobulins with a new salt-promoted adsorbent.
abstract: In this work a highly acetylated-ethylenediamine-Novarose (HA-EDA-Novarose) gel was synthesized and used as a new agarose-based salt-promoted adsorption chromatography (SPAC) matrix to effectively isolate serum immunoglobulins without the need of denaturing conditions. Samples of human serum in 0.5 M Na2SO4, 10 mM 3-(N-morpholino)-propane-sulfonic acid (MOPS), pH 7.6 were applied to a chromatographic column packed with the SPAC gel. Immunoglobulins (Igs) with affinity for the HA-EDA ligands were specifically adsorbed to the matrix, non-bound serum proteins were readily removed by washing the column with the same feed solution buffer. Bound Igs were effectively and very gently eluted by simply removing the salt from the feed solution buffer. The elution buffer consisted thus of only 10 mM MOPS, at pH 7.6 and no salt. The salt-dependent adsorption capacity of this system was estimated to be 7.3 mg/ml with protein recovery of about 93%. Sodium dodecyl sulfate-polyacrylamide gel (SDS-PAGE) electrophoresis analysis, radial immunodiffusion and enzyme-linked immunosorbent assays showed that immunoglobulins G, M and A (IgG, IgM and IgA) were the main components present in the elution fraction. The new SPAC adsorbent was used to purify Igs from human serum and IgG and IgA from non-pure commercially available Igs preparations in a very gentle single step.
authors.affiliation: Departamento de Ciencias Químico Biológicas, Universidad de Sonora, Rosales y Blvd., Luis Encinas S/N, Hermosillo, Sonora 83000, Mexico.
authors.forename: Maria
authors.initials: M
authors.lastname: del Carmen Candia-Plata
authors.forename: Javier
authors.initials: J
authors.lastname: García
authors.forename: Roberto
authors.initials: R
authors.lastname: Guzmán
authors.forename: Jerker
authors.initials: J
authors.lastname: Porath
authors.forename: Luz
authors.initials: L
authors.lastname: Vázquez-Moreno
mesh_terms: D000327:Adsorption; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007136:Immunoglobulins / Q000097:blood / Q000302:isolation & purification*; D012492:Salts / Q000737:chemistry*
chemical_list: D007136:Immunoglobulins; D012492:Salts