Automated method for quantification of 20 amino acids in cell culture media during biopharmaceutical development.
Biotechniques
; 76(1): 27-36, 2024 01.
Article
en En
| MEDLINE
| ID: mdl-37997819
ABSTRACT
Herein, 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.
Palabras clave
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Productos Biológicos
/
Aminoácidos
Límite:
Animals
Idioma:
En
Revista:
Biotechniques
Año:
2024
Tipo del documento:
Article
País de afiliación:
India