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Protein A-G-L & Affinity Conjugates

BioVision offers a wide range of Agarose, Sepharose & Magnetic beads coupled to Protein A, Protein G, Protein L, Protein A/G, Protein A/G/L or Jacalin to purify monoclonal and polyclonal antibodies from culture media, serum, ascites fluid or hybridoma supernatants and to isolate antibody/antigen complexes in immunoprecipitation experiments.

Proteins A, G and L have been widely used for IgG purification. BioVision’s Protein A, G and L resins consist of purified recombinant protein A, G & L respectively that have been covalently coupled to 6% cross-linked Agarose/Sepharose beads. The recombinant protein A and G contain only IgG binding domains and are purified from E. coli via multiple chromatographic steps (not affinity purified with human IgG). The albumin binding domain, cell wall binding domain and other non-specific regions have been removed to ensure the maximum specific IgG binding. Unlike Protein A and Protein G, which bind primarily through Fc regions (i.e., heavy chain) of immunoglobulin, Protein L binds Igs through interaction with the light chains. The recombinant Protein L contains only Kappa light chain binding region and thus has the ability to bind a wider range of Ig classes (IgG, IgM, IgA, IgE and IgD) and subclasses. Single chain variable fragments (ScFv) and Fab fragments also bind to Protein L. Purity of these recombinant proteins is > 98% by SDS-PAGE and HPLC. Endotoxin level is < 0.1 EU/mg of Protein. The 6-His-tag on N-terminus can be used for affinity purification or detection using anti-His-tag antibodies. BioVision has performed extensive research to prepare special Hi-Bind™ protein A & Hi-Bind™ protein G-Agarose beads that give a higher binding capacity for IgG, higher flow rate & minimal leaching of the ligand than the other regular Protein A & Protein G-Agarose on the market. The Protein A, G and L resins display high chemical and physical stability as well as show high flow rate, hydrophilicity & high gel strength. These resins are available in several convenient pack sizes and can be regenerated and reused multiple times when stored properly.

BioVision’s high-quality Jacalin Sepharose exhibits specific, high-yield purification of human IgA from serum or colostrum and thus can also be used for removing contaminating IgA from IgG.

Relevance/Importance:
  • Purification of broad range of monoclonal and polyclonal antibodies.
  • Isolation and purification of IgG from serum, culture media, ascites fluid or hybridoma supernatants.
  • Isolation and purification of antigens and immune complexes by immune co-precipitation.
  • Depletion of immunoglobulins from serum, culture media, ascites fluid or hybridoma supernatants.
Key Features:

Enhanced Specificity
• BioVision Recombinant Protein A and Protein G contain only IgG-binding regions of native Protein A and native Protein G respectively. The cell wall binding region, albumin binding region and other non-specific regions have been eliminated to ensure the maximum specific IgG binding.

High Binding Capacity and High Flow Rate
• Lower molecular weight of the affinity protein permits higher density of its immobilization and elevated capacity.


Minimal leaching of the ligand
• Our conjugation chemistry ensures stable immobilization that prevents loss of immobilized affinity proteins during elution and regeneration of the beads.

Versatility
• Binds a broad range of IgG species and subclasses.

Durability
• Can be regenerated and reused multiple times without deterioration of performance.

Convenience
• Available in multiple sizes to match all your needs.



Figure: Purification of IgG using Protein A Sepharose (Cat. # 6501): (a) SEC analyses of IgG purified from rabbit serum. 1 ml rabbit serum in Protein A IgG Binding Buffer was loaded on 1 ml Protein A Sepharose & IgG was purified according to the protocol. 200 µl of starting sample (A) (serum) and purified IgG (B) were analyzed on Superdex 200 HR 10/30 column at 0.5 ml/min in 50 mM Tris, 0.25 M NaCl pH 7.5. (b) SDS-PAGE (10%) of purified IgG under reduced conditions. Lane 1: Marker, Lane 2: IgG fraction (5 µg).