Antibody purification

affinity chromatography - protein A and protein G Sepharose.

Ana Grodzki, Elsa Berenstein

Research output: Contribution to journalArticle

36 Citations (Scopus)

Abstract

Affinity chromatography relies on the reversible interaction between a protein and a specific ligand immobilized in a chromatographic matrix. The sample is applied under conditions that favor specific binding to the ligand as the result of electrostatic and hydrophobic interactions, van der Waals' forces and/or hydrogen bonding. After washing away the unbound material the bound protein is recovered by changing the buffer conditions to those that favor desorption. The technique has been used not only to isolate antigen-specific antibodies but also to remove specific contaminants from biological samples. Methods are described for the purification of immunoglobulins, namely IgG, IgG fragments and subclasses, using the high affinity of protein A and protein G coupled to agarose. In the Subheading 3 there are also protocols for affinity purification using a specific ligand coupled to commercial matrices like CNBr- Sepharose 4-B and Affigel.

Original languageEnglish (US)
Pages (from-to)33-41
Number of pages9
JournalMethods in molecular biology (Clifton, N.J.)
Volume588
StatePublished - Mar 4 2010
Externally publishedYes

Fingerprint

Antibody Affinity
Staphylococcal Protein A
Affinity Chromatography
Sepharose
Ligands
Immunoglobulin G
Proteins
Hydrogen Bonding
Static Electricity
Hydrophobic and Hydrophilic Interactions
Immunoglobulins
Buffers
Antigens
Antibodies

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Antibody purification : affinity chromatography - protein A and protein G Sepharose. / Grodzki, Ana; Berenstein, Elsa.

In: Methods in molecular biology (Clifton, N.J.), Vol. 588, 04.03.2010, p. 33-41.

Research output: Contribution to journalArticle

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