Product – Byzen Pro

Forget acid elution and improve you antibody purification

Byzen Pro® is an affinity chromatography resin in which a novel Protein A mutant is used as a ligand. The product provides a new antibody purification scheme where antibody is eluted under neutral pH by changing temperature, avoiding risk of aggregation of antibody.

volume item code
5 ml 0809N05
25 ml 0809N25

Product Information

Human antibody is purified from cell culture by using Protein A affinity chromatography. Conventional Protein A resin uses strongly acidic buffer to elute antibody, exposing the target antibody to risk of aggregation and decreased activity. In contrast, our temperature-responding Protein A (Byzen Pro®) allows antibody to be eluted under neutral pH by simply changing temperature. This eliminates any risk of aggregation and decreased activity of antibody, improving manufacturing efficiency of antibody drugs.

Antibody is eluted under neutral pH.
Byzen Pro® needs no acidic conditions. No risk of aggregation of antibody.
Binding and elution are controlled by temperature.
Antibody is bound below 10ºC and eluted between room temperature to 40ºC.
A buffer of your choice can be used for elution.
PBS buffer can be used for the Protein A step. Buffer exchange won’t be necessary.
Good antibody specificity
Byzen Pro® is as specific as the conventional Protein A, showing excellent impurity elimination capability.
A recovery ratio is 95% or more.

1. PURIFICATION WITH A CLOSED COLUMN

graph
CAUTION: Column pressure should stay below 0.3 MPa when operating chromatography equipment or a pump.

Column Volume
1.0 ml
Bed Height
2.5 cm
Buffer
20 mM Phosphate, 150 mM NaCl, pH 8.0
Flow Rate
1.0 ml/min
Binding temperature
4 ºC
Elution temperature
40 ºC
Sample
Human IgG (1 mg) + cell lysate (1 ml)

PROCEDURE

  1. Attach a 5 mL loop before the column, and wash the column with 5 CV of H2O.
  2. Immerse the column and the loop into a 4℃ water bath and then equilibrate the column with buffer.
  3. Wait for 5 min for temperature equilibration, and then inject a sample.
  4. After wash the column to remove impurities, stop the pump. Take the column and the loop out of the 4ºC water bath and immerse them into a 40ºC water bath.
  5. Wait for 5 min for temperature equilibration, and then start the pump to elute antibody.

2. PURIFICATION WITH OPEN COLUMN (NO JACKET)

Purification may be performed by placing the column in a cold room/refrigerator or an incubator set at desired temperature. It is recommended to calibrate a time required to equilibrate the column before use.
When a flow rate by gravity is too slow, use an aspirator, a pump or a syringe to withdraw buffer from the column.

Buffer
20 mM Phosphate, 150 mM NaCl, pH 8.0
Flow Rate
Gravity
Binding temperature
4 ºC
Elution temperature
40 ºC

PROCEDURE

  1. Pack the column with Byzen Pro® using a pipette (a bed height of 2 cm or more is recommended when the concentration of antibody is low).
  2. Steps 3) and 4) should be performed in a cold room or by placing the column in a refrigerator set at 4ºC.
  3. Equilibrate the column with 5 CV of buffer by gravity (buffer should be pre-cooled at 4ºC).
  4. Make sure that the column temperature is 4ºC, and gently apply a sample onto the surface of the resin to allow the sample to flow by gravity.
  5. Apply a 4ºC buffer onto the surface of the resin without disturbing the surface to wash the column. Repeat this step until all impurities are removed.
  6. Steps 7) and 8) should be performed by placing the column in an incubator set at 40ºC.
  7. Make sure that the column temperature is 40ºC, and gently apply a 40ºC buffer onto the surface of the resin without disturbing the surface to elute antibody by gravity.

STORAGE

  1. Wash the column with 5 CV of H2O.
  2. For a closed column, replace water with 20% ethanol and store at 4ºC.
  3. For an open column, replace water with 20% ethanol. Take the resin out of the column and store in a closed container at 4ºC.

SPECIFICATION

Bead Structure Cross-linked polyvinyl alcohol
Mean Particle Size 70 μm
Max. Back Pressure 0.3 Mpa、3bar
Flow Rate (Recommend) 125 – 250 cm/h
pH Range (Recommend) pH 6 – pH 8
Binding Temperature 4ºC
Eluting Temperature 37 ºC – 40 ºC
Dynamic Binding Capacity 26 – 34 mg/ml (20 cm column)
Storage Condition 20% ethanol at 4ºC

Specificity

  • Human IgG1, IgG2 and IgG4
  • Rabbit IgG and Goat IgG