Automated method for quantification of  20 amino acids in cell culture media during biopharmaceutical development

Biotechniques. 2023 Nov 24. doi: 10.2144/btn-2023-0068. Online ahead of print.ABSTRACTHerein, a step-by-step protocol for simultaneous detection of 20 amino acids commonly present in cell culture media is described. The protocol facilitates detection of both primary and secondary amino acids through a two-step precolumn derivatization strategy using ortho-phthalaldehyde and 9-fluorenylmethyl chloroformate as derivatizing agents. The separation of derivatized amino acids with varying hydrophobicity is achieved through reverse-phase chromatography. The amino acids are simultaneously detected in a single workflow through the use of Variable Wavelength Detector at 338 and 262 nm. The protocol is applicable for both mammalian and bacterial cell culture matrices with an option for automation of precolumn derivatization.PMID:37997819 | DOI:10.2144/btn-2023-0068

Source: BioTechniques - November 24, 2023 Category: Biotechnology Authors: Ankita Dubey Srishti Joshi Kratika Upadhyay Ansuman Mahato Anurag S Rathore Source Type: research