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Extracellular Glucose Depletion as an Indirect Measure of Glucose Uptake in Cells and Tissues Ex Vivo
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Evaluating Dextrose-Derived Interference in Low-Concentration Monoclonal Antibody Quantification for Clinical In-Use

Rakesh Dachineni1, Nida Naseer2, Matthew Myers2

  • 1Drug Product Development, BioCMC, Teva Branded Pharmaceutical Products R&D LLC, West Chester, Pennsylvania, 19380, USA.

AAPS Pharmscitech
|March 6, 2026
PubMed
Summary

Glucose degradation products in dextrose 5% in water (D5W) IV bags interfere with UV absorbance protein quantification. Protein-A affinity chromatography offers a reliable method for accurate monoclonal antibody concentration measurement in D5W.

Keywords:
5-Hydroxymethylfurfural (5-HMF)UV absorbanceaffinity chromatographyintravenous infusion bagsmass spectroscopy

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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.