Chromolith® WP 300 Protein A – Fast monoclonal antibody quantitation

Affinity chromatography is a most selective technique which takes advantage of very specific molecular interactions, for example antigen and antibody. The Chromolith® WP 300 Protein A HPLC column is designed to monitor monoclonal antibody titer and yield determination from cell-culture supernatants. Analytical scale procedure helps to optimize the titer of monoclonal antibody for the optimal time for harvest of the monoclonal antibody products. Chromolith® WP 300 Protein A column could be used for separation of all IgGs (except class 3). Columns provides extremely fast separations and could be used longer minimizing.

Separation of monoclonal antibodies

Constant binding efficiency at any flow rate

  • High-speed separation at high flow rate due to excellent mass transfer properties of the monolithic skeleton
  • Separation of IgG demonstrates the extreme time savings and high separation efficiency made possible with Chromolith® protein A columns.
  • IgG was well separated with excellent peak symmetry
  • At 5 mL/min the total analysis time is less than 1 minute and the net column backpressure is only 21 bar
  • Antibody binding is not affected by flow rate

Monolithic column provides low column backpressure at high flow rates

Monolithic column provides low column backpressure at high flow rates

 

Column separation reproducibility and life time

Reproducibility

Chromatographic overlay of 50 injections

For mAb titer determination, it is necessary to use a column offering a high reproducibility. The Chromolith® protein A column offers highly reproducible elution of bound antibodies with constant retention time and low change in peak area.

Chromatographic overlay of 50 injections


Stability test

Chromolith® WP 300 Protein A column offers high stability and reproducibility

More than 53.000 column volumes of 100 mM sodium phosphate buffer including 10.000 pH shifts were applied to the column.
 

Stability test

 

Batch-to-batch reproducibility

Batch-to-batch reproducibility

 

Materials Table

Product Modification I.d. Length Type Content Product No.
Chromolith® WP 300 Protein A Protein A 4.6 mm 25 mm   1 HPLC column 152258
Chromolith® WP 300 RP18 RP-18 4.6 mm 100 mm   1 HPLC column 152270
Chromolith® WP 300 RP18 RP-18 4.6 mm 50 mm   1 HPLC column 152271
Chromolith® WP 300 RP18 Guard Cartridge RP-18 4.6 mm 10 mm b 3 guard cartridges 152272
Chromolith® WP 300 RP18 Guard Cartridge RP-18 4.6 mm 5 mm b 3 guard cartridges 152273
Chromolith® WP 300 RP8 RP-8 4.6 mm 100 mm   1 HPLC column 152265
Chromolith® WP 300 RP8 RP-8 4.6 mm 50 mm   1 HPLC column 152266
Chromolith® WP 300 RP8 Guard Cartridge RP-8 4.6 mm 10 mm b 3 guard cartridges 152267
Chromolith® WP 300 RP8 Guard Cartridge RP-8 4.6 mm 5 mm b 3 guard cartridges 152268
Chromolith® WP 300 RP4 RP-4 4.6 mm 100 mm   1 HPLC column 152260
Chromolith® WP 300 RP4 RP-4 4.6 mm 50 mm   1 HPLC column 152261
Chromolith® WP 300 RP4 Guard Cartridge RP-4 4.6 mm 10 mm b 3 guard cartridges 152262
Chromolith® WP 300 RP4 Guard Cartridge RP-4 4.6 mm 5 mm b 3 guard cartridges 152263
Chromolith® WP 300 Epoxy Epoxy 4.6 mm 100 mm   1 HPLC column 152250
Chromolith® WP 300 Epoxy Epoxy 4.6 mm 50 mm   1 HPLC column 152251
Chromolith® WP 300 Epoxy Epoxy 4.6 mm 25 mm   1 HPLC column 152252
Chromolith® WP 300 Epoxy Guard Cartridge Epoxy 4.6 mm 10 mm b 3 guard cartridges 152253
Chromolith® WP 300 Epoxy Guard Cartridge Epoxy 4.6 mm 5 mm b 3 guard cartridges 152254