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Protein adsorption-desorption on electrospray capillary walls--no influence on aggregate distribution
Suvajyoti Guha1, Joshua R Wayment, Mingdong Li
1Departments of Mechanical Engineering, Chemistry and Biochemistry, University of Maryland, College Park, MD 20742, USA.
Journal of Colloid and Interface Science
|April 24, 2012
Summary
Protein adsorption in glass capillaries does not affect aggregate size distribution during electrospray (ES). Gelatin passivation effectively reduces antibody adsorption, ensuring reliable protein aggregate analysis.
Area of Science:
- Analytical Chemistry
- Biophysics
- Materials Science
Background:
- Protein adsorption on capillary surfaces can lead to desorption, potentially altering aggregate size distributions.
- Accurate quantification of protein aggregates is crucial in biopharmaceutical development and quality control.
Purpose of the Study:
- To investigate the impact of protein adsorption and desorption on protein aggregate size distribution.
- To evaluate the effectiveness of different surface passivation methods for glass capillaries used in electrospray applications.
Main Methods:
- Differential Mobility Analysis (DMA) was employed to measure the size distribution of proteins eluting from bare and passivated glass capillaries.
- Surface passivation was performed using gelatin and Bovine Serum Albumin (BSA).
Main Results:
- No significant differences in aggregate size distributions were observed between unpassivated and passivated capillaries at steady state.
- Gelatin passivation demonstrated superior efficacy in reducing adsorption of Rituxan and polyclonal human IgG compared to BSA.
- Gelatin passivation remained stable for several days and across a pH range of 4.8-9.0.
Conclusions:
- Desorbing proteins from capillary walls do not significantly influence protein aggregate distribution measurements.
- Gelatin passivation is a robust and effective method for minimizing protein adsorption in glass capillaries for electrospray analysis.
- The findings support the use of gelatin-passivated capillaries for reliable protein aggregate quantification.
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