生物制药开发过程中细胞培养基中20种氨基酸的自动定量方法。

IF 2.2 4区工程技术 Q3 BIOCHEMICAL RESEARCH METHODS

BioTechniques Pub Date : 2024-01-01 Epub Date: 2023-11-24 DOI:10.2144/btn-2023-0068

Ankita Dubey, Srishti Joshi, Kratika Upadhyay, Ansuman Mahato, Anurag S Rathore

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引用次数: 0

摘要

本文描述了用于同时检测通常存在于细胞培养基中的20种氨基酸的分步方案。该方案通过使用邻苯二醛和9-氟酰氯甲酸甲酯作为衍生剂的两步柱前衍生化策略，促进了一级和二级氨基酸的检测。通过反相色谱法分离具有不同疏水性的衍生氨基酸。通过使用338和262 nm的可变波长检测器，在单个工作流程中同时检测氨基酸。该方案适用于哺乳动物和细菌细胞培养基质，可选择柱前衍生化自动化。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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

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.

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来源期刊

BioTechniques 工程技术-生化研究方法

CiteScore

4.40

自引率

0.00%

发文量

审稿时长

3.3 months

期刊介绍： BioTechniques is a peer-reviewed, open-access journal dedicated to publishing original laboratory methods, related technical and software tools, and methods-oriented review articles that are of broad interest to professional life scientists, as well as to scientists from other disciplines (e.g., chemistry, physics, computer science, plant and agricultural science and climate science) interested in life science applications for their technologies. Since 1983, BioTechniques has been a leading peer-reviewed journal for methods-related research. The journal considers: Reports describing innovative new methods, platforms and software, substantive modifications to existing methods, or innovative applications of existing methods, techniques & tools to new models or scientific questions Descriptions of technical tools that facilitate the design or performance of experiments or data analysis, such as software and simple laboratory devices Surveys of technical approaches related to broad fields of research Reviews discussing advancements in techniques and methods related to broad fields of research Letters to the Editor and Expert Opinions highlighting interesting observations or cautionary tales concerning experimental design, methodology or analysis.