Overview
Harlan Laboratories can purify your monoclonal and polyclonal
antibodies using a wide a variety of purification methods, based on
a minimum starting volume of 10 ml. The correct choice of
purification method will depend on a number of variables. These
include the use for which the antibodies are intended, the species
in which it was raised, its class and subclass, and the source
(e.g. supernatant, serum, or ascites). We are happy to discuss your
purification options and develop the purification procedure that
provides the best balance between yield, purity, and cost. A full
profile with analysis by SDS page and production records is
supplied with each purification. Antibody purification methods
available include:
- Ammonium Sulphate Precipitation
- Protein A Affinity Chromatography
- Protein G Affinity Chromatography
- Ion Exchange Chromatography (anion and cation)
- Ig Affinity
- Antigen Affinity
- Size Exclusion Chromatography
Ammonium Sulphate
Precipitation
- This Method is useful for concentration and partial
purification of antibodies from most sources and all species.
Although, on its own the yields of antibody are impure, it is the
method of choice when combined with ion exchange for large volumes
of anti-sera. It is recommended for antibody purification of tissue
culture supernatant.
Protein
A
- This process binds the Fc region of major sub-classes of mouse
and rat IgG to varying degrees. The results with protein A are
usually high purity and high yields. It is useful for large-scale
purifications of tissue culture supernatants.
Protein
G
- Protein G can be used for purifying antibody from most sources.
Like protein A, the results are usually of high purity and high
yield. It is applicable to a wider range of immunoglobulins than
protein A, but elution procedures can be much harsher.
Ion Exchange
Chromatography
- This method is used for all sources of antibodies. It is
applicable to all IgG isotypes. It is usually the method of choice
when purifying antibodies from ascites, as the monoclonal can be
resolved from the host immunoglobulins.
Ig Affinity
- This method can be used to isolate mouse IgM and IgG3, and for
isolating monoclonal antibodies from supernatants which contain
high levels of serum Ig.
Antigen
Affinity
- This is the method of choice when specific antibody is required
from polyclonal anti-sera. It requires pure antigen for the best
results and can sometimes result in inactive antibody.
Size
Exclusion
- This method is appropriate for IgM antibodies from all sources.
It is used when IgM antibodies need to be separated from polyclonal
anti-sera. It is not recommended as a single stage.
When purification applies to your antibody protocol, this
information need to be filled out in the
Antibody Services Production Questionnaire.
Contact us at antibodies@harlan.com for
pricing and additional information.