Isolation of human serum immunoglobulins with a new salt-promoted adsorbent

Maria del Carmen Candia-Plata, Javier García, Roberto Z Guzman, Jerker Porath, Luz Vázquez-Moreno

Research output: Contribution to journalArticle

4 Citations (Scopus)

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.

Original languageEnglish (US)
Pages (from-to)211-217
Number of pages7
JournalJournal of Chromatography A
Volume1118
Issue number2
DOIs
StatePublished - Jun 23 2006

Fingerprint

Adsorbents
Immunoglobulins
Salts
Adsorption
Chromatography
Serum
Immunoglobulin A
Morpholinos
Propane
Buffers
ethylenediamine
Sulfonic Acids
Immunoglobulin G
Immunoglobulin M
Gels
Immunosorbents
Immunodiffusion
Electrophoresis
Washing
Sodium Dodecyl Sulfate

Keywords

  • Agarose-based adsorbents
  • HA-EDA
  • IgA
  • IgG
  • IgM
  • Immunoglobulins
  • Salt-promoted adsorption chromatography

ASJC Scopus subject areas

  • Analytical Chemistry

Cite this

Isolation of human serum immunoglobulins with a new salt-promoted adsorbent. / Candia-Plata, Maria del Carmen; García, Javier; Guzman, Roberto Z; Porath, Jerker; Vázquez-Moreno, Luz.

In: Journal of Chromatography A, Vol. 1118, No. 2, 23.06.2006, p. 211-217.

Research output: Contribution to journalArticle

Candia-Plata, Maria del Carmen ; García, Javier ; Guzman, Roberto Z ; Porath, Jerker ; Vázquez-Moreno, Luz. / Isolation of human serum immunoglobulins with a new salt-promoted adsorbent. In: Journal of Chromatography A. 2006 ; Vol. 1118, No. 2. pp. 211-217.
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