Monoclonal antibodies are widely used in the diagnosis, prevention and treatment of diseases. Since the first monoclonal antibody drug was launched in 1986, nearly 40 kinds of McAb drugs have been listed, and more than 300 kinds of McAb drugs have been in clinical study. McAb drugs have become one of the fastest-growing drug fields in the world.

In the production process of monoclonal antibody, the cost of McAb drugs can account for 50-70% of its production cost for that this process requires rigorously for the efficiency and cost. Protein A affinity medium is the most critical material for antibody separation and purification as its properties and cost determine the purity, yield and cost of McAb products.

Currently, majority of antibodies IgG molecules use Protein A affinity chromatography for purification. Protein A is the surface protein of cell wall found in staphylococcus aureus with a molecular weight of 42 kDa, and its five different binding regions can bind IgG Fc fragments with a strong specific affinity. Each Protein A molecule can bind at least two antibody IgG molecules.

Protein A Tanrose 4FF

Protein A Tanrose 4FF is the universal affinity chromatography medium for the separation and purification of monoclonal antibodies, polyclonal antibodies, or Fc- fusion labels. Protein A has five IgG binding regions and other regions with unknown functions. Recombinant protein A removes binding sites with albumin on cell surface and contains only five IgG binding regions, reducing nonspecific adsorption.

Based on highly cross-linked 4% agarose gel, Protein A Tanrose 4FF can be purified at a relatively high flow rate by monoclonal antibody and polyclonal antibody, which is generally accepted by industrial customers.

NameProtein A Tanrose 4FFProtein A Solid
Bead structure6% highly cross-linked agaroseHighly rigid agarose
Particle size45-165µm45-120µm
Mean particle size90μm75µm
Binding capacity>20mg(humanIgG)/ml(media)>28mg(humanIgG)/ml(media)
pH stability*3-9((long term)2-11(short term)3-10((long term)2-12(short term)
Chemical stability*8 M urea, 20% ethanol, 6 M guanidine hydrochloride (8 days), stable at pH 2-9.
Max. flow rate250cm/h500cm/h
Operating pressure≤0.3MPa≤0.5MPa
Storage solution 20% ethanol 20% ethanol
Storage temp.4-8℃4-8℃

Buffer Preparation

The water and buffer are recommended to be filtered with 0.22 μm or 0.45 μm filter membrane before use.

Equilibrium liquid/eluate : 20 mM Na₂HPO₄, 0.15M NaCl, pH 7.0;

Eluent: 0.1 M glycine, pH 3.0;

Neutralizer: 1 M Tris-HCl, pH 8.5.

Sample Purification

After packing the Protein A Tanrose 4FF, various conventional medium and low pressure chromatographic systems can be used.

  • Fill the pump pipe with deionized water. Remove the plug, and connect the column to the chromatographic system. Open the lower outlet, then connect the pre-column to the chromatographic system and tighten it.
  • Flush storage buffer with deionized water of 3-5 times column volume.
  • Use at least 5 times the column bed volume of equilibrium liquid to equilibrate the column.
  • Use pump or sample ring to load samples. Note: the increase of the viscosity of the sample will lead to the large back pressure of the column, even if the volume of the sample is very small. The injection volume should not exceed the binding capacity of the column, as a large sample volume may also cause the large back pressure, making the injector more difficult to use.
  • Flush the column with eluate until UV absorption reaches a stable baseline (generally at least 10-15 times of the column volume).
  • Use 5-10 times column volume of eluent, collect target protein components. The elution components need to be neutralized immediately. It is generally recommended to use neutralizer of 1/10 volume of the elution component.

Protein A Tanrose 4FF

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