Instrumental analytical techniques for quality assessment of biotechnology- derived monoclonal antibodies

Abstract

This thesis consists of four parts: introduction, experimental, results and discussion, General conclusion in addition to 68 figures, 54 tables, 135 references and the Arabic summary. Part (I): General introduction and literature review This part includes a general introduction to biopharmaceuticals, their production, their quality control procedures and their stability. Introduction to the structure and production of monoclonal antibodies, in addition to their types and the revolution it caused in the therapeutic field. A general background about development and regulatory pathways of biosimilars and interchangeability were summarized. Anti-TNF products structures and clinical uses and a brief description about their structural characterization were presented. A comprehensive review of different methods of analysis reported in literature for the determination of the studied products was discussed in this part as well. Part (II): Experimental This section includes methods development, optimization, calibration and their application to antibodies formulations. Size exclusion high performance liquid chromatography was developed and validated for the determination of the studied products in the presence of their high molecular weight aggregates. Reversed phase high performance liquid chromatography assay was used for determination of oxidized variants. Tapestation electrophoresis was carried out to interpret the nature of aggregate. Dynamic light scattering was used a complementary assay with size exclusion high performance liquid chromatography in evaluating high molecular weight aggregates and low molecular weight fragments.