Temperature and pH-dependent behaviors of mAb drugs: a case study for Trastuzumab

Sert, Fatma and Hız, Defne and Gülmez, Mert and Cankurtaran, Selen Ezgi and Kayalan, Cemre Irmak and Kurt, Hasan and Yüce, Meral (2022) Temperature and pH-dependent behaviors of mAb drugs: a case study for Trastuzumab. Scientia Pharmaceutica, 90 (1). ISSN 0036-8709 (Print) 2218-0532 (Online)

Full text not available from this repository. (Request a copy)


The distortions in the high-order structure of therapeutic monoclonal antibodies (mAbs) under different environmental conditions acutely affect mAb stability, resulting in altered safety, efficacy, and shelf-life profiles. The overall stability of mAbs depends on many factors, and it re-quires complementary techniques for an in-depth analysis. The stability of mAbs can be character-ized by differential centrifugal sedimentation (DCS), differential scanning calorimetry (DSC), differential scanning fluorimetry (DSF), and size exclusion chromatography (SEC) techniques. In this report, temperature-ramped dynamic light scattering (DLS), and circular dichroism (CD) spectroscopy were employed as complementary tools to show how temperature and pH affect the aggregation of a model mAb, trastuzumab, in solution. The results showed that the aggregation onset temperature of trastuzumab defined by DLS was 75 °C, which decreases the amount of β-sheets and causes a slight increase in helix structures. Moreover, the melting temperature of trastuzumab was determined to be between 80–83 °C by temperature-ramped CD spectrophotometry, which is in line with the Tm of trastuzumab’s Fab region tested with DSC. Thus, unfolding and aggregation of trastuzumab start simultaneously at 75 °C, and unfolding triggers the aggregation. The tempera-ture-ramped CD and DLS methods are robust tools to determine the thermal behavior of biosimilars in various solution conditions. Their complementary usage provides solid scientific background for regulatory applications and a better understanding of mAb instability and its relationship with structural changes.
Item Type: Article
Uncontrolled Keywords: biophysical characteriza-tion; colloidal stability; complementary tools; conformational stability; thermal stability
Divisions: Faculty of Engineering and Natural Sciences
Sabancı University Nanotechnology Research and Application Center
Depositing User: Fatma Sert
Date Deposited: 22 Aug 2022 21:59
Last Modified: 22 Aug 2022 21:59
URI: https://research.sabanciuniv.edu/id/eprint/44124

Actions (login required)

View Item
View Item