Pierce IgG Purification Buffers have been optimized to provide the highest efficiency of IgG binding and elution for most species. The use of other buffer formulations may significantly alter not only the binding capacity, but the volumes of wash buffer needed to ensure a good purification. When used with Pierce IgG Purification Buffers, chromatography on Pierce Immobilized Protein A and Protein A/G columns will result in maximal binding. It is the buffer composition, not the pH, that optimizes immunoglobulin binding to Protein A and Protein A/G.

The binding capacities of Immobilized Protein A, Immobilized Protein G, and Immobilized Protein A/G with serum of different species show improved binding using our Buffers. The Protein A IgG Binding Buffer (Product #’s 21001, 21007 available below) and the IgG Elution Buffer (Product #’s 21004, 21009) are designed for high yield isolation of mouse monoclonal IgG1. They have long shelf lives and are premixed for maximum ease of use.

We also offer a Gentle Ag/Ab Binding Buffer (Product #21012) and Gentle Ag/Ab Elution Buffer (Product #21013). These products were designed to eliminate your worries about acidic elutions and the time-consuming step of neutralizing your fractions. The Gentle Buffers can usually dissociate the antigen-antibody interaction efficiently under neutral pH conditions. This system can also usually dissociate the interaction between Protein A and IgG effectively. Our researchers have tested the effect of exposure to Gentle Elution Buffer on monoclonal antibody activity. The following murine monoclonal anti-HIV reagents were used for the experiment: anti-RT, anti-120N and anti-env41. The monoclonals were incubated overnight in Gentle Elution Buffer, then desalted. The isolated fractions then were analyzed in an ELISA system for any loss of binding activity. All three monoclonals retained full antigen binding capability. The following figure illustrates the purification of mouse IgG using the Gentle Ag/Ab Buffers. To see the figure, click on the chart button. 

Highlights

• Assures maximum recovery of IgG from immobilized Protein A, G and A/G
• Perfect for binding and elution of antibody from antigen
• Premixed and easy to use
• Specially formulated and prefiltered
• Eliminated use of harsh acidic conditions (Gentle Ag/Ab Buffers)
• Specially formulated for neutral elutions (Gentle Ag/Ab Buffers)

Binding Capacities with Different Buffers Expressed as mg IgG Bound per 2 ml of gel

Purification of Mouse IgG using Gentle Ag/Ab Buffer System.
Purification was with a 1 ml column of Immobilized Protein A and the absorbance of each 5 ml fraction was measured.