Rapid one-step purification of goat immunoglobulins by immobilized metal ion affinity chromatography

Valerie Boden, Joy Winzerling, Mookambeswaran Vijayalakshmi, Jerker Porath

Research output: Contribution to journalArticle

59 Citations (Scopus)

Abstract

A rapid, single step purification of immunoglobulins from goat serum was achieved using immobilized metal ion affinity chromatography (IMAC) on a new high capacity gel, Novarose, coupled to tris(2-aminoethyl)amine (TREN) chelated with copper. When goat serum was adsorbed to this gel in buffer pH 7 at 11 cm/h (8.6 ml/h), the immunoglobulin fraction was recovered in a decreasing linear pH gradient at about pH 5.5. When the adsorption buffer was adjusted to pH 6.0 and the linear velocity increased to 110 cm/h (221 ml/h), an immunoglobulin fraction of greater than 95% homogeneity was obtained. Protein purity was assessed by silver-stained native and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS PAGE). The capacity of the gel for immunoglobulins was 17 mg immunoglobulin/ml at the low flow rate with adsorption at pH 7 and 15 mg immunoglobulin/ml at the high flow rate with adsorption at pH 6. No problems of back pressure or gel compression were observed at the higher linear velocity. The mild elution pH, high flow rate, and synthetic nature of the ligand support make this new metal-chelating gel a powerful alternative to the use of other currently available commercial gels commonly used for immunoglobulin purification.

Original languageEnglish (US)
Pages (from-to)225-232
Number of pages8
JournalJournal of Immunological Methods
Volume181
Issue number2
StatePublished - Apr 26 1995

Fingerprint

Affinity Chromatography
Goats
Immunoglobulins
Metals
Ions
Gels
Adsorption
Buffers
Proton-Motive Force
Serum
Silver
Sodium Dodecyl Sulfate
Copper
Polyacrylamide Gel Electrophoresis
Ligands
Pressure
Proteins

Keywords

  • Immobilized metal ion affinity chromatography
  • Immunoglobulin purification
  • Tris(2-aminoethyl)amine

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology
  • Biotechnology

Cite this

Rapid one-step purification of goat immunoglobulins by immobilized metal ion affinity chromatography. / Boden, Valerie; Winzerling, Joy; Vijayalakshmi, Mookambeswaran; Porath, Jerker.

In: Journal of Immunological Methods, Vol. 181, No. 2, 26.04.1995, p. 225-232.

Research output: Contribution to journalArticle

Boden, Valerie ; Winzerling, Joy ; Vijayalakshmi, Mookambeswaran ; Porath, Jerker. / Rapid one-step purification of goat immunoglobulins by immobilized metal ion affinity chromatography. In: Journal of Immunological Methods. 1995 ; Vol. 181, No. 2. pp. 225-232.
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