Antibody purification

ammonium sulfate fractionation or gel filtration.

Ana Grodzki, Elsa Berenstein

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

Antibodies can be purified by a variety of methods based on their unique physical and chemical properties such as size, solubility, charge, hydrophobicity and binding affinity. This chapter focuses on ammonium sulfate precipitation as a convenient first step in antibody purification in that, it allows the concentration of the starting material and the precipitation of the desired protein. The principle of ammonium sulfate precipitation lies in "salting out" proteins from the solution. The proteins are prevented to form hydrogen bonds with water and the salt facilitates their interaction with each other forming aggregates that afterward precipitate out of solution. Gel filtration or size- exclusion chromatography is also discussed in this chapter. Gel filtration is based on the relative size of protein molecules and it is of great value to separate IgMs, exchange buffers and/or desalt solutions. The columns designed to separate the proteins are composed of porous beads and the proteins will flow through the packed column inside and around the beads, depending on its size.

Original languageEnglish (US)
Pages (from-to)15-26
Number of pages12
JournalMethods in molecular biology (Clifton, N.J.)
Volume588
StatePublished - Mar 4 2010
Externally publishedYes

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Ammonium Sulfate
Gel Chromatography
Antibodies
Proteins
Hydrophobic and Hydrophilic Interactions
Solubility
Hydrogen
Buffers
Salts
Water

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Antibody purification : ammonium sulfate fractionation or gel filtration. / Grodzki, Ana; Berenstein, Elsa.

In: Methods in molecular biology (Clifton, N.J.), Vol. 588, 04.03.2010, p. 15-26.

Research output: Contribution to journalArticle

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