Evaluating Dextrose-Derived Interference in Low-Concentration Monoclonal Antibody Quantification for Clinical In-Use...

Area of Science:

• Biochemistry
• Analytical Chemistry
• Pharmaceutical Development

Background:

• Monoclonal antibody (mAb) drug product development requires stability and compatibility studies.
• 5% dextrose in water (D5W) is a common intravenous diluent posing analytical challenges.
• Steam sterilization of D5W generates glucose degradation products (GDPs) that interfere with UV absorbance at 280 nm (A280).

Purpose of the Study:

• To investigate the impact of GDPs on A280-based protein quantification of mAbs diluted in D5W.
• To evaluate the influence of temperature and storage duration on GDP accumulation.
• To identify and implement a reliable method for accurate mAb quantification in D5W.

Main Methods:

• Investigated GDP formation in D5W IV bags under various storage conditions.
• Assessed the interference of D5W with A280-based quantification of a model mAb at concentrations ≤ 1.0 mg/mL.
• Developed and validated a Protein-A affinity chromatography method for mAb quantification.

Main Results:

• GDP levels in D5W varied and increased with prolonged storage at elevated temperatures.
• D5W significantly interfered with A280-based mAb quantification, with persistent variability even after baseline correction.
• Protein-A affinity chromatography accurately quantified mAbs (0.01–1.0 mg/mL with UV, 0.005–1.0 mg/mL with fluorescence detection) in D5W.

Conclusions:

• D5W-derived GDPs introduce significant discrepancies in A280-based protein measurements.
• Protein-A affinity chromatography effectively mitigates GDP interference, providing accurate mAb quantification in D5W.
• This method is crucial for reliable in-use studies of mAb drug products diluted in D5W.