Evolocumab is a fully human monoclonal antibody designed for the treatment of hyperlipidemia by Amgen. Evolocumab is a human IgG2 monoclonal antibody that targets the proprotein convertase subtilisin/kexin type 9 (PCSK9). PCSK9 is a protein that targets LDL receptors for degradation, therefore reducing the liver's ability to remove LDL-cholesterol (LDL-C), or "bad" cholesterol, from the blood. This protein heterogeneity results from sequence variations generated from proteolysis or transcriptional/translational errors, from post-translational modifications (PTMs), and from degradation of products which are formed during processing or final product storage. Therefore, detailed characterization and sensitive analytical techniques are required to ensure the safety, quality and efficacy of therapeutic protein products.

The EU regulation and guidelines of biosimilar have been enacted. However, the cutting-edge protein characterization method can serve as a technical barrier. This study was performed to prepare and establish standard operating procedure (SOP) for improve analytical techniques of Evolocumab by Liquid chromatography -mass spectrometry-based analytical method.