1.Details

BG ProG FF antibody purification Resins specifically binds to the Fc region of antibodies through coupled Protein G ligand, achieving high-selectivity adsorption of antibody molecules and Fc fusion proteins. After elution under acidic conditions, high-purity target products can be collected. Target High-purity antibodies can be obtained from samples such as ascites, serum, and culture medium through a single step of affinity chromatography process. BG ProG FF antibody purification medium uses agarose gel as the matrix and recombinant Protein G as the ligand, offering high binding capacity, excellent physicochemical stability, low ligand detachment, and long lifetime. 

2.Features

Chromatographic operation Procedure

The Antibody Purification Resins (GP-ProA) is widely used for the separation and purification of various antibodies. Chromatographic operations typically include column packing, equilibration, ample loading, washing, elution, and regeneration steps.

Equilibration: Equilibrate the chromatography column with 5-10 column volumes (CV) of equilibration buffer (20 mM PB + 0.15 M NaCl, pH 7.0, with appropriate concentration of NaCl to inhibit non-specific adsorption) until the conductivity and pH of the effluent remain constant (consistent with the equilibration buffer).

Sample Loading: The sample buffer should be as consistent as possible with the equilibration buffer. Solid samples can be dissolved and prepared in equilibration buffer; dilute sample solutions can be dialyzed with equilibration buffer; concentrated sample solutions can be diluted with equilibration buffer. To avoid column clogging, samples should be centrifuged or microfiltered (0.45 μm) before loading. The sample loading volume should be determined based on the resins binding capacity and the target protein concentration in the sample.

Washing: After sample loading, continue washing with equilibration buffer until the baseline is reached.

Elution: Elute with elution buffer (20 mM sodium acetate, pH 3.0-4.0 or 0.1 M glycine, pH 3.0), and collect the effluent. After elution, immediately neutralize the collected antibody solution to neutral pH with alkaline buffer (such as 1 M Tris/HCl, pH 9.0) to maintain antibody biological activity and prevent inactivation.

Regeneration and Cleaning in place: After several uses (5-10 cycles, depending on the type and source of raw materials and experimental requirements), the medium requires regeneration and cleaning in place.(1) Wash the column with 0.1 M acetic acid or 20% ethanol for 3-5 CV, then wash to neutrality with buffer for reuse;(2) Alternatively, wash the column with 0.05 M NaOH + 1 M NaCl or 6 M guanidine hydrochloride for 3-5 CV, rinse with 3-10 CV of pure water, then wash to neutrality with buffer for reuse.

Other considerations: During column packing, use, and storage, avoid allowing the column to run dry and prevent air bubbles from entering.
3.Specification

 

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