The Shimadzu PPSQ-51A/53A N-Terminal Protein Sequencer automates Edman reaction, LC separation, detection and data analysis to determine amino acid sequence from the N-terminal of proteins . The system offers various features, such as low running cost thanks to the reduced consumption of analytical reagents, easy operation and enhanced data processing functions including chromatogram subtraction from cycle to cycle. Though mass spectrometry has been used in the identification of amino acids, Edman degradation offers many distinct advantages- guaranteed N-terminal sequence of proteins, no sample pre-treatment required, differentiation of isobaric amino acids (leucine and isoleucine), identification possible even from unknown proteins not registered in data bases.
Isocratic separation ensures stable baselines and provides higher sensitivity analysis of PTH-amino acids. Retention times are also highly reproducible which allows confident peak identification between successive cycles.
Specialized protein sequencer software makes it simple to perform the reprocessing of chromatograms, theoverlay of multiple chromatograms, and the automatic estimation of amino acid sequences, which are requiredfor sequence analysis.Sequences are automatically estimated after analysis ofeach cycle is complete. Up to four candidate amino acids are displayed together with their certainty levels.
LabSolutions PPSQ software provides compliance with FDA 21 CFR Part 11 guidelines and enables compliance with the security,user management and audit trail requirements specified by FDA 21 CFR Part 11.
The Guidelines on biopharmaceutical molecules (ICH-Q6B) says “Terminal amino acid analysis is performed to identify the nature homogeneity of the amino- and carboxyl-terminal amino acids’’. N-terminal amino acid analysis is conducted to confirm the type and uniformity of N-terminal amino acids. This requires measurement of the relative quantities of molecular variants by an appropriate analytical method, when the targeted biopharmaceutical molecules is non-uniform with respect to the terminal amino acid. In addition, EMA (European Medicines Agency) published the guideline on biosimilars, “Guideline on similar biological medicinal products” in May, 2014. The Shimadzu Protein Sequencer 51A/53A serves to fill this key requirement through the automated sequencing of the N-terminal amino acids .