Separations

ABT offers a selection of Protein A, A/G, G and L ligands and formats to purify total or specific antibodies from serum, ascites fluid, or tissue culture supernatant.

Protein A

rProtein A consists of a single polypeptide chain which contains five highly homologous antibody-binding domains. The binding site is located on the Fc region of immunoglobulin and rProtein A has affinity for IgG from a variety of mammalian species and for some IgA and IgM. The recombinant Protein A shares identical binding properties to IgG as the Cowan I strain of natural Protein A. 

Protein G

Protein G is a well-established ligand for affinity chromatography resins used for isolation and characterization of immunoglobulins. Protein G binds to the Fc region of mammalian IgGs thus exhibiting high affinity for many IgG subclasses including human, rabbit, mouse, rat, sheep, pig, horse, cow or goat. In contrast to protein A, protein G binds all human and mouse IgG subclasses, including human IgG3 and mouse IgG1 as well as all rat IgG subclasses. Protein G also binds to CH1 portion of the Fab region making this ligand well suited for purification of more complex constructs such as bispecific antibodies. 

Download the full Protein G Benchmarking Study that compares three commercially available protein G resins - and see how ABT's Protein G Agarose 4 Rapid Run came out on top!

Download the ABT Antibody Purification Resins brochure to see the full range of Antibody Purification Resins for Protein A, A/G, G and L

Affinity Chromatography is a technique that separates tagged proteins and other biomolecules using biological interactions. This technique has high selectivity and is widely used to obtain proteins with high purity at high yields. ABT manufactures a variety of activated beads used for His-tag, Antibody, GST and Avidin/Streptavidin Purifications.

His-Tag Purification

His-tagged proteins can be purified by a single-step affinity chromatography, using immobilized metal ion affinity chromatography (IMAC). A number of formats are commercially available with Ni-NTA matrices being the most widely used.

ABT offers a range of IMAC resins and pre-packed cartridges for purification of His-tag proteins. ABT's Nickel NTA Agarose Cartridges 5 ml are pre-packed ready to use products for fitting into the MPLC, FPLC and AKTA™ devices. They can be purchased in units of 1 or 5 cartridges packed with 5 ml of Nickel NTA Agarose Resin.

Download the His-Tag Purification Application Guide to find out more about the range of resins and cartridges available for His-Tag Purification

REFERENCES:

• Martínez-Fleta, Pedro et al. “SARS-CoV-2 Cysteine-like Protease Antibodies Can Be Detected in Serum and Saliva of COVID-19–Seropositive Individuals" J Immunol November 4, 2020.

 

Ion Exchange Chromatography (IEX) separates biomolecules on the basis of their net surface charge. Biomolecules will provide different interaction properties depending on the net charge, density and surface charge distribution. Biomolecules without a net charge at a pH equivalent to their isoelectric point (PI) will not interact with the resin. Depending on the working pH, the biomolecule will exhibit interaction to anion or cation exchanger media. Therefore, at a pH above its pI, the biomolecule will interact with a positively charged resin (anion exchanger) and at a pH below its pI, the biomolecule will show affinity to a negative charged resin (cation exchanger).

ABT offers a wide range of plain and crosslinked agarose beads with different agarose concentrations (2, 4, 6, 8, & 10%) in different particle distributions: Standard (50-150µm), Fine(20-50µm), and Macro (150-350µm). ABT also has developed Rapid Run™ which are based on highly crosslinked 4% and 6% agarose matrices used for industrial scale separations.

Sepadextran™ (25 & 50) is a beaded gel filtration medium prepared by crosslinking dextran. Both types are available in three different particle sizes Medium, Fine, and Superfine in which ABT supplies in its dry form.

ABT offers a great range of pre-activated resins that are designed to couple ligands via stable & uncharged covalent linkages that minimize leakage of the affinity ligand and reduce non-specific binding. Two different chemistries of pre-activated beads are available: Glyoxal and Aminoethyl beads. The choice of Glyoxal or Aminoethyl will depend on the ligand to be immobilized, the accessibility of the reactive groups and the direction/orientation required for the binding to the support.

Glyoxal Agarose

ABT offers a family of preactivated agarose base matrices specifically developed for the preparation of affinity chromatography resins. These base matrices are preactivated with aldehydes groups and are marketed under the trade name Glyoxal Agarose (GA). Glyoxal chemistry enables the coupling of proteinaceous ligands via reductive amination of the primary amino groups on the ligand surface. This chemistry enables stable conjugation of ligands via multipoint covalent attachment and has been proven advantageous over other immobilization chemistries due to the increased stabilization of the coupled ligand.

Download the Glyoxal Agarose App Note to find out more about the advantages of these preactivated supports

Agarose Bead Technologies are committed to provide quality products along with responsive service, designed to meet and exceed the needs and customer expectations. ABT's quality systems comply with the international standards for Quality Management Systems as defined by the ISO 9001:2015.