MegaPoly AT antibody affinity analytical column is the newly launched high pressure liquid chromatography column by Bestchrom.

The column is prepacked with MegaPoly AT resin, which is a polystyrene divinylbenzene-based resin with average particle size of 20μm. Its alkali resistant Protein A ligands are suitable for the rapid measuring and small-size preparation of bio-products including monoclonal antibodies and Fc fusion protein. On account of the convenience, efficiency and reliability of the method, the product is widely applicable in the cell culture condition screening in R&D as well as the quality control in production.

Major applications

• Culture condition screening: Select the cell lines and their culture condition by expressed titer detection.

• Purification quality control: Expressed volume determination of antibody or Fc fusion protein from upstream feed liquid.

• Content determination of antibody or Fc fusion protein after downstream purification.

• Small scale antibody preparation

Products features

• High resolution, high pressure resistance: average particle size of 20μm, max pressure is 20 MPa, reliable and robust.

• Convenient and rapid operation: single sample testing can be done within several minutes.

• Recombinant alkali resistant ligands: better binding capacity and wider application, could withstand CIP with 0.1-0.5M NaOH

Table 1. Technical parameters of MegaPoly AT antibody affinity analytical column.

Standard sample testing

Standard curve of human IgG sample

Experiment condition:

Column: MegaPoly AT analytical column, 2.1mmD × 30mmH

 A similar product from one imported brand, column volume 0.1mL

System: Agilent 1200 HPLC, UV Monitor: 280nm

Equilibration buffer (Buffer A): 50mMPB + 150mM NaCl pH 7.0

Elution buffer (Buffer B): 100mM acetic acid pH 2.5

Flow rate: 3mL/min

Sample: human IgG

Sample load volume: 20μL IgG standard sample

Concentration: 0.125mg/mL—20mg/mL

Standard curve of one bi-specific antibody

Experimental condition:

Column: MegaPoly AT analytical column, 2.1mmD × 30mmH 

System: Agilent 1200 HPLC, UV Monitor: 280nm

Equilibration buffer (Buffer A): 50mM PB  + 150mM NaCl pH 7.0

Elution buffer (Buffer B): 100mM phosphate PH 2.5

Flow rate: 1mL/min

Sample: bi-specific antibody, original sample concentration 5.04mg/mL

Sample load volume: 20μL

Bi-specific antibody concentration: 0.5mg/mL—5mg/mL

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Standard curve of Fc fusion protein

Experiment condition:

Column: MegaPoly AT analytical column, 2.1mmD × 30mmH

System: Agilent 1200 HPLC, UV monitor: 280nm

Equilibration buffer (Buffer A): 50mM PB + 150mM NaCl pH 7.0

Elution buffer (Buffer B): 100mM phosphate PH 2.5

Flow rate: 1mL/min

Sample: Fc fusion protein, original concentration 11.6mg/mL

Sample load volume: 20μL

Fc fusion protein concentration: 1mg/mL—10mg/mL

?Standard curve of human IgG sample under different elution conditions

Experiment condition:

Column: MegaPoly AT analytical column, 2.1mmD × 30mmH

System: Agilent 1200 HPLC, UV monitor: 280nm

Equilibration buffer (Buffer A): 50mM PB + 150mM NaCl pH 7.0

Elution buffer (Buffer B): one from the following list

•100 mM citric acid, pH 2.5

•100 mM acetate, pH 2.5

•100 mM phosphate, pH 2.5

•100 mM glycine, pH 2.5

•12 mM HCl, pH 1.9

Flow rate: 3mL/min

Sample: Human IgG 

Sample load volume: 20μL IgG standard sample

Concentration: 0.125mg/mL—20mg/mL

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