Abstract
INTRODUCTION.
Medicines, based on recombinant DNA technology, are widely used in the treatment of cancer, immune-inflammatory, and infectious diseases. Research on the development of new protein-derived drugs, including monoclonal antibodies, as well as methods for quality control of such drugs, is ever increasing. Peptide mapping allows it to verify the primary protein structure, genetic stability, and identify structural changes.
AIM.
Systematization of current methodological approaches to the development of peptide mapping methods.
DISCUSSION.
One of the main methods for confirming protein authenticity is peptide mapping based on enzymatic hydrolysis of protein to produce a unique set of peptide fragments. Despite the uniqueness of each method, they all share common principles of sample preparation and analysis, as well as regulatory requirements. Method development is a complex, multi-step process. Currently, various enzymes are used for protein cleavage, but trypsin remains the “gold standard”. Ready-made solutions for protein cleavage reactions are becoming increasingly common such as highly specific and reproducible kits, sample preparation approaches based on immobilized enzymes on magnetic particles, and automated procedures. These options minimize errors in manual sample preparation, improve the reproducibility of results, and reduce analysis time.
CONCLUSIONS.
Current peptide mapping methodology is evolving towards increased reproducibility and efficiency through the introduction of standardized and automated sample preparation solutions while maintaining trypsin as the primary enzyme for protein cleavage. Information on approaches used will enable researchers to navigate the diversity of available analytical solutions, accelerate method development, and ensure reliable quality control of protein-based medicines.