In monoclonal antibody (mAb) manufacturing, accurate antibody titre determination and monitoring are crucial for clonal selection, optimisation of expression rates, process control and scale-up. For accurate antibody titre quantification, Protein-A Affinity HPLC offers excellent reproducibility, robustness and precision over ELISA-based assays.
The SOLAD™ Protein-A is a novel affinity HPLC column, based on the next-gen SOLAD™ platform technology, for quick separation and robust quantification of various antibodies. It provides rapid, accurate titre analysis with linearity across a wide concentration range, without any carryover.
SEM of SOLAD Particles (2500x magnified)
❖ Wide dynamic range
❖ Fast Analysis
❖ High sensitivity & specificity for mAbs
❖ Long lifetime
SEM of SOLAD Particles (10000x magnified)
SOLAD™ is a proprietary product range of highly monodisperse, spherical, durable and reproducible non-porous silica particles manufactured by Glantreo. For protein and antibody analysis, SOLAD™ offers excellent mechanical and chemical stability, with minimal fouling and non-specific adsorption, compared to porous silica and polymeric particles.
The SOLAD™ Protein-A column combines a 10µm sized SOLAD particle with recombinant Protein A ligand through proprietary cutting-edge bonding technology. The result being a hydrophilic resin surface with a high pH stability and a rugged and reproducible column. The column offers superior efficiency and resolution, without analyte carryover.
The SOLAD™ Protein-A column can accurately measure mAb titres at various phases of development due to its high loading capacity and excellent repeatability. This column is also suitable for small-scale purification of mAbs for initial characterisation due to its reduced Protein A leaching. The column is available in PEEK and Bioinert Stainless Steel column hardware in various dimensions.
Technical Specifications and Applications of SOLAD Protein-A Column
Rapid separation of IgG from Impurities (Fig. 1)
Column: 4mm x 50 mm PEEK, Binding buffer (A): 50 mM Sodium Phosphate pH 7.4, Elution buffer (B): 50 mM Sodium Phosphate pH 2.5, Gradient: 0%B: 0-0.5 min, 100%B: 0.5-1.1 min, 0%B: 1.1-4 min, Flow rate: 2 mL/min, Detection: UV @ 280 nm, Temperature: 25 C, Sample: 20 µL cell culture supernatant containing mAb
Figure 1 depicts a 20 μL injection of mAb feedstock onto the SOLADTM Protein A column. The proprietary column technology yields sharp, concentrated peaks and fast, efficient elution.
Wide Dynamic Range (Fig. 2)
As can be seen in Figure 2, SOLADTM Protein-A delivers accurate titre measurement with linearity, with a dynamic loading capacity in excess of 300 µg.
Flexible Flow Rates (Fig. 3)
The SOLADTM Protein-A column can be used from flow rates upto 2.5 mL/min, as seen in Figure 3. This allows fast separation of antibodies without loss of efficiency.