mAb titer for process development (or purification of small amounts of mAb for further characterization fast separation).

• High throughput clonal selection (60 seconds per sample)
• Robust mAb recovery at 97%
• Analytical separation of all IgG subclasses (human and mouse), Protein G offers alternate selectivity for those IgG molecules that do not bind to Protein A
• Flow-rate independent separations
• No diffusion, no pores and no void volume make transport between mobile and stationary phase very rapid
• Extremely fast separations speed up method development time and decrease costs
• Key applications are quantitative determination of IgG (fermentation titer calculation)

Affinity chromatography is one of the most effective ways to determinate the titer of monoclonal antibodies. Protein A and Protein G affinity chromatography columns absorb the IgG molecules onto the affinity resins which contain either immobilized protein A ligand or protein G ligand, then the remaining impurities and byproducts from the fermentation broth can be removed. Elution of the purified monoclonal antibody and quantification by comparing the peak area to a calibration curve allows rapid measurement of the protein concentration.

Agilent Bio-Monolith Protein A and Protein G affinity columns are designed for analytical separation of all IgG subclasses (human and mouse). They deliver fast and accurate quantitation of mAb titer (sub 1 minute) and purification of small amounts of mAb for subsequent CQA analysis by another complementary technique, such as Aggregate Analysis or Charge Variant Analysis, and can easily be combined into a 2D-LC workflow. They are compatible with HPLC and UHPLC systems, including the Agilent 1100, 1200, 1260, and 1290 Bio-inert Quaternary LC Systems.