Aggregation Determination using nano|Filter Vials® by SEC
The aggregation of protein therapeutics has become a major concern for the pharmaceutical industry and regulatory agencies. Protein aggregates can cause an adverse immune response and are typically monitored throughout the formulation and production of bio-therapeutics. Monitoring aggregates helps to minimize risks from therapeutic proteins in clinical applications by optimizing early formulations to reduce aggregation during production, storage, handling, and shipping.
Antibodies are clarified using Thomson nano|Filter Vials®, 0.2 µm PES Membrane (Part#: 15535-200 or 500) and analyzed for purity using SEC (Size Exclusion Chromatography). 10 µL of purified antibody was placed into the nano|Filter Vials® and 2 µL was injected onto the HPLC. Fig. 1 & 2 show a chromatogram of mAb1 with low abundance multimers. Fig. 3 & 4 show a chromatogram of mAb2 with dissociated antibody fragments.
Method & Sample Preparation
- Agilent 1260 Infinity HPLC
- Column: 4.6x300 mm Bio-SEC3
- Buffer: 1x PBS (Phosphate Buffered Saline)
- Flow Rate: 0.2 mL/min
- Injection: 2 µL
- 10 µL of purified antibody
- Thomson nano|Filter Vials®, 0.2 µm PES Membrane
fig.1Chromatogram of Antibody mAb1
fig.2Chromatogram of Antibody mAb2
fig.3Zoomed in version of the chromatogram in Fig. 2 of Antibody mAb1 to better visualize low abundance dimer.
fig.4Zoomed in version of the chromatogram in Fig. 3 of Antibody mAb2 to better visualize aggregate peak.
Thomson nano|Filter Vials®, 0.2 µm PES Membrane Part#: 15535-200 (qty 200) | 15535-500 (qty 500)
Aggregation Determination using nano|FV™ by SEC
The aggregation of protein therapeutics has become a major concern for the pharmaceutical industry and regulatory agencies. ... Antibodies are clarified using Thomson nano|Filter Vials® ... , and analyzed for purity using SECPDF Application