Aggregation Determination using nano|Filter Vials® by SEC

Introduction

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.

prothena nanoFV elements

Method & Sample Preparation

Method

  • 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

Sample Prep

  • 10 µL of purified antibody
  • Thomson nano|Filter Vials®, 0.2 µm PES Membrane

Chromatograms

prothena nanoFV elements1

fig.1Chromatogram of Antibody mAb1

prothena nanoFV elements2

fig.2Chromatogram of Antibody mAb2

prothena nanoFV elements3

fig.3Zoomed in version of the chromatogram in Fig. 2 of Antibody mAb1 to better visualize low abundance dimer.

prothena nanoFV elements4

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)

Title

Aggregation Determination using nano|FV™ by SEC

Description

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 SEC

PDF Application

Product Used in Application

nano|Filter Vial® - PES 0.2µm

Non-Split Septum, Grey Cap

pn#15535-200

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