Qualitative and quantitative examination of non-specific protein adsorption on filter membrane disks of a commercially available high throughput chromatography device.

Abstract: High throughput process development (HTPD) platforms provide time and resource effective solutions for optimization of resin as well as membrane based chromatography processes. They enable successful implementation of Quality by Design by allowing us to investigate more variables and wider ranges than would be possible using traditional lab-scale based experiments. This paper presents an investigation about non-specific protein adsorption that was found to occur on the membrane disks used in a commercially available high throughput device. Both qualitative as well as quantitative analytical tools were used in this investigation. Two biotherapeutic protein molecules, granulocyte colony stimulating factor (GCSF) and Transtuzumab were selected as model proteins in this study. Qualitative assessment was performed using Attenuated Total Reflectance Fourier-Transform Infrared Spectroscopy (ATR-FTIR), Scanning Electron Microscopy (SEM) and Atomic Force Microscopy (AFM). An empirical model based on contact angle measurement was developed for quantitative assessment of the amount of adsorbed protein. The results indicated that while significant non-specific adsorption was observed in case of highly hydrophobic GCSF molecule, adsorption was minimal in the case of Transtuzumab. The paper proposes a novel use of contact angle measurement for quantitative estimation of adsorbed protein. The study also highlights the importance of accounting for non-specific adsorption when using these HTPD devices. [Copyright &y& Elsevier]