By Chromatography

Group Members
Protein purification is a series of processes intended to
isolate a single type of protein from a complex
mixture.

Separation of one protein from all others is typically

the most laborious aspect of protein purification.

Separate the protein and non-protein parts of the

mixture, and finally separate the desired protein from
all other proteins.
Pepsin

Urease
Protein size
Physico-chemical properties
Binding affinity
Biological activity
Preparative Purification
like Enzymes

Analytical Purification
like Protein structure ,
modification
Extraction

Precipitation

Ultracentrifugation
Repeated freezing
Thawing
Homogenization by high pressure
Filtration
 Flow of the solution containing the protein
through a column packed with various
materials.
Different proteins interact differently with the
column material, and can thus be separated by
the time required to pass the column.
Size exclusion chromatography
Ion exchange chromatography
HPLC
Size-exclusion chromatography (SEC) is a
chromatographic method in which molecules
in solution are separated by their size, and in
some cases molecular weight
It is usually applied to large molecules or
macromolecular complexes such as proteins
and industrial polymers.

A size exclusion column

Gel filtration is used in fractionation mode,
uses porous particles to separate multiple
components in a sample on the basis of
differences in their size.
Mixture of similar charged ions separated by using ion
exchange resin .
Reversible exchange of similar charged ions.
Cations or Anions can be separated PRINCIPLE
Reversible exchange of ions b/w ions present in the
solution & ion exchange resins.
HPLC is a form of liquid chromatography used to
separate compounds that are dissolved in
solution.
Compounds are separated by injecting a sample
mixture onto the column.