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Antibody Purification Services
Antibody Purification Services

Antibody Affinity Purification Services

Bio-Synthesis provides large scale custom antibody purification services with a wide array of service options. The correct choice of purification method depends upon the class/subclass of the antibody, the species in which it was raised and the intended use of the antibodies. We will purify antibodies from bioreactors, supernatants, ascites, and serum from small (ml) to large volume (liters) purification using protein A or protein G affinity chromatography, as well as rare ones such as protein L. Depending on the application of the antibody, purity and recovery will likely determine the strategy of purification. All antibody purifications include a confirmation check using SDS-PAGE.

Our staff will be happy to discuss your antibody purification options and develop the type of purification that will provide the best balance between yield, purity and cost. Our Antibody Purification Service Options provide clients with solutions they can trust.

BSI supplies an Antibody Purification Information Sheet, with each inquiry to assist in meeting customer specifications.

Antibody Purification Options

The following is a list of our current antibody purification services. If you are in need of special purification, large quantities or you do not see a package that suits your needs, please feel free to contact us for a quote.

Description Cat No.
Protein A/G Affinity Purification • up to 100 ml of sera or ascites • QC by SDS-PAGE IMMCHEM-PURE-001
Protein A/G Affinity Purification • 100-1,000 ml of sera or ascites • QC by SDS-PAGE IMMCHEM-PURE-002
Protein A/G Affinity Purification • up to 2.0 liters of culture supernatant • QC by SDS-PAGE IMMCHEM-PURE-003
Protein A/G Affinity Purification • > 2.0 liters of culture supernatant • QC by SDS-PAGE IMMCHEM-PURE-004
Protein L Affinity Purification • up to 100 ml of sera or ascites • QC by SDS-PAGE IMMCHEM-PURE-005
Protein L Affinity Purification •100-1,000 ml of sera or ascites • QC by SDS-PAGE IMMCHEM-PURE-006
Peptide Specific Affinity Purification • up to 10 ml serum or ascites • include ELISA assay on crude and purified sample • QC by SDS-PAGE and peptide coupling to column IMMCHEM-PURE-007
Peptide Specific Affinity Purification • 20 ml serum or ascites • include ELISA assay on crude and purified sample • QC by SDS-PAGE and peptide coupling to column IMMCHEM-PURE-008
Peptide Specific Affinity Purification • up to 100 ml serum or ascites • include ELISA assay on crude and purified sample • QC by SDS-PAGE and peptide coupling to column IMMCHEM-PURE-009
Peptide Specific Affinity Purification • 100-1,000 ml serum or ascites • include ELISA assay on crude and purified sample • QC by SDS-PAGE and peptide coupling to column IMMCHEM-PURE-010
Ammonium Sulfate Precipitation Egg Yolks • up to 10 yolks IMMCHEM-PURE-011
Egg IgY Affinity Purification • 20 ml egg yolks (~2 egg yolks) IMMCHEM-PURE-012
Egg IgY Affinity Purification • up to 100 ml egg yolks (~10 egg yolks) IMMCHEM-PURE-013
Antigen Specific Chicken IgY Affinity Purification • from 20 ml egg yolks (~ 2 egg yolks) IMMCHEM-PURE-014
Antigen Specific Chicken IgY Affinity Purification • up to 100 ml egg yolks (~10 egg yolks) IMMCHEM-PURE-015
Protein-based Immunoaffinity Purification • up to 100 ml of antisera IMMCHEM-PURE-016
GST/His/Trx/Flag Affinity Column Cross-Absorption IMMCHEM-PURE-017
Ion Exchange Purification • up to 100 ml of sera or ascites IMMCHEM-PURE-018
Ion Exchange Purification • 100-1,000 ml of sera or ascites IMMCHEM-PURE-019
Low EU (Endotoxin Unit) Protein A/G Purification • up to 100 ml of sera or ascites IMMCHEM-PURE-020
Low EU Protein A/G Purification • 100-1,000 ml of sera or ascites IMMCHEM-PURE-021
Low EU Protein A/G Affinity Purification • up to 2.0 liter of culture supernatant IMMCHEM-PURE-022
Low EU Protein A/G Affinity Purification • > 2.0 liter culture supernatant IMMCHEM-PURE-023
Low EU Ion Exchange Purification • up to 100 ml of sera or ascites IMMCHEM-PURE-024
Low EU Ion Exchange Purification • 100-1,000 ml sera or ascites IMMCHEM-PURE-025
Low EU Protein L Purification • up to 100 ml of sera or ascites IMMCHEM-PURE-026
Low EU Protein L Purification • 100-1,000 ml of sera or ascites IMMCHEM-PURE-027

Antibody Purification Methods

Antibody purification can be divided into two groups: precipitation methods and chromatographic methods. The latter is grouped further into non-affinity and affinity chromatography.

The choice of purification methods available include:

  • Polyethylene Glycol (PEG) and /or Ammonium Sulphate Precipitation
  • Immunoaffinity purification
  • Protein A affinity Chromatography
  • Protein G Affinity Chromatography
  • Protein A/G affinity chromatography
  • Ig Antibody Affinity Purification
  • Antibody Purification by Ion exchange chromatography
  • Hydrophobic interaction chromatography
  • Size exclusion chromatography
  • Ammonium sulfate precipitation
  • HPLC purification
  • Procedures and methodology used for the purification and Quality Control report (including analysis of yield, purity and immunoreactivity) are provided with each purified product

Antibody purification using Protein A, G, or A/G

Protein A/G is a recombinant or genetically engineered protein that combines both Protein A and Protein G binding properties. Protein A/G is designed to contain four Fc binding domains from Protein A and two from Protein G. Protein A or Protein G purification removes the IgG fraction based on the specificity of these proteins for the Fc portion of the IgG. Protein A is produced from Staphylococcus aureus. It has the capacity to bind at least two molecules of IgG. The binding is specific to the Fc portion and does not affect the antigen binding sites. Protein G is isolated from Group G streptococci and binds the Fc region of the IgG in a similar manner to Protein A. Protein A and Protein G have differing binding efficiencies for IgG from different species. It is important to check this before deciding which method to use. For example, Protein G works well with sheep Ig, but Protein A does not. Neither Protein A nor Protein G will bind to chicken Ig. Care should be taken when eluting the antibody from the column to avoid denaturation of the antibody.

Type of Sample Isotype Average Concentration
Serum lgG Serum: 1 to 3 mg/ml
Ascitic Fluid Ascitic fluid: 1 to 5 mg/ml
Cell culture supernatant Cell Culture supernatant : 5 to 20 ug/ml
Service Includes:
  • Preparation of protein A or G affinity column, loading a dilution of bound antibodies
  • SDS-PAGE analysis of purified antibodies

Peptide Immunoaffinity Purification

The most commonly used method to purify antigen-specific antibodies from crude sera is immunoaffinity purification. Unlike Protein A or G, the non-specific Ig fraction is not retained. In this procedure, peptide antigen is bound covalently to a solid support. The antibodies within the polyclonal sample that are specific for the peptide antigen bind to the support column. The unbound antibodies are removed from the column by washing and the specific antibodies are eluted from the column. The product of immunoaffinity purification is highly specific antibodies. Immunoaffinity purification can occasionally cause denaturation of the antibody due to the conditions used to elute the bound antibody from the column. It is important to compare the response generated by the purified sample against the response generated by the crude sera.


Type of Sample Min Peptide Quantity Average Concentration
Serum 5 to 7 mgs 50 to 100 ug of immuno-purified antibodies/mL
Service Includes:
  • Peptide immobilization on sepharose beads
  • Loading of antiserum on the affinity column and elution of bound antibodies
  • ELSIA testing on crude serum and immunopurified antibodies

IgY Purification from Egg Yolk

Chicken antibodies IgY are the equivalent to mammalian IgG antibodies and can be isolated from serum or egg yolk. The egg yolks contain at least the same concentration of IgY antibodies found in chicken or rabbit serum. IgY can be used in most biochemical and cellular applications, including Western Blots. As the Fc region of chicken IgY is sufficiently different from mammalian IgG chicken antibodies do not cross-react with mammalian proteins, and do not bind with mammalian rheumatoid factors, Fc-receptors or proteins A or G. Therefore, the background signal or false response in certain immunochemical assays is reduced.

Type of Sample Yield of Purification
Eggs, or egg yolks ~10 to 15 mg of lgY / egg yolk
Service Includes:
  • Preparation of the egg yolk extract by two-step precipitation
  • Loading of the egg yolk extract on the ion exchange column and elution of bound antibodies
  • ELSIA testing on starting material and immunopurified antibodies
  • SDS-PAGE analysis of purified antibodies

Polyethylene Glycol and /or Ammonium Sulphate Precipitation

This method is useful for concentration and partial purification of antibodies from most sources and all species including purification of IgY's from chicken egg yolks. Although on its own the yields of antibodies are impure, it is the method of choice when combined with ion exchange for large volumes of antisera and or followed by Antigen Affinity purification to enrich for specific antibodies. It is not recommended for purification of tissue culture supernatant.

Ig Antibody Affinity Purification

This method can be used to isolate monoclonal antibodies from supernatants which contain high levels of serum Ig. It is the choice for secondary antibody production.

Size Exclusion/Gel Filtration Chromatography

A single gel matrix or a sequence of size exclusion columns can be used for purification of IgM and IgG antibodies. The purification strategy employed is dependent on the immunoglobulin class and the source of the material.

Antibody Purification by Ion Exchange Chromatography

This is usually the method of choice when purifying antibodies from ascites, as the monoclonal can be resolved from the host immunoglobulins. It has been used in adjunct to ammonium sulfate precipitation for purifying antibodies from ascites fluid or where Protein A and Protein G elution conditions may damage the antibody. It is also applicable to all IgG isotypes.

Aseptic/Low Endotoxin Removal, Purification and Testing

Purification under aseptic conditions in a dedicated, limited access facility to generate clean, low-endotoxin antibody preparations.


Reference/Citing:
  1. Different prelamin A forms accumulate in human fibroblasts: a study in experimental models and progeria
  2. 1,25-DIHYDROXYVITAMIN D3-INDUCED GENES IN OSTEOBLASTS: UNCOVERING NEW FUNCTIONS FOR MENINGIOMA 1 AND SEMAPHORIN 3B IN SKELETAL PHYSIOLOGY
  3. Fetal Death in Mice Lacking 5α - Reductase Type 1 Caused by Estrogen Excess
  4. A splicing variant of a death domain protein that is regulated by a mitogen-activated kinase is a substrate for c-Jun N-terminal kinase in the human central nervous system

Visit Literature Vault for more citing on Custom Polyclonal Antibodies

Ordering & Contact Information

  • Please contact us for additional information or send an email to info@biosyn.com.
  • You may also request an online quote.
  • To contact us by phone, please call 1-972-420-8505 or Fax at 1-972-420-0442
  • Orders may be placed using a purchase order (PO) or by credit card through our secure online ordering system.
  • When using a credit card ( creditcard ) it will be billed under "Bio-Synthesis, Inc."