HIGH THROUGHPUT PREPARATION OF AMINO ACIDS USING WATERS TECHNOLOGIES ACCQ-TAG ULTRA AUTOMATION KIT AND CELL CULTURE STANDARDS

Significance of the Topic

The accurate quantification of amino acids in cell culture media is essential for monitoring nutrient levels, optimizing cell growth, and ensuring consistent biopharmaceutical production. Automating this process reduces manual errors, enhances throughput, and supports reproducible data generation.

Objectives and Study Overview

This study aimed to develop an automation-compatible AccQ-Tag Ultra reagent kit and standardized scripts for three liquid handling platforms (Andrew+, Hamilton Microlab STAR, Tecan Freedom EVO 100/4). The goal was to compare automated preparation against manual workflows and assess performance in terms of precision, accuracy, and linearity for 26 amino acids.

Methodology

• Sample Preparation: 96-well format derivatization using AccQ-Tag Ultra reagents with norvaline internal standard.
• Automation Platforms: Scripts optimized for Andrew+, Hamilton STAR, Tecan EVO with minimal user intervention and barcode integration.
• Analytical Conditions: Waters ACQUITY UPLC H-Class system with TUV detection at 260 nm, AccQTag Ultra column (2.1 × 100 mm, 1.7 µm), flow rate 0.7 mL/min, column temperature 43 °C, injection volume 1 µL.

Instrumentation

• Waters ACQUITY UPLC H-Class and H-Class Bio with TUV detector.
• Automated liquid handlers: Andrew+ workstation, Hamilton Microlab STAR, Tecan Freedom EVO 100/4.
• Empower 3 software for data processing and Excel for statistical evaluation.

Main Results and Discussion

• Precision: Automated methods achieved mean CV ≤3.9% across platforms, matching manual preparation.
• Accuracy: All amino acids showed <15% deviation in automated runs on Hamilton and Tecan systems.
• Linearity: Calibration curves exhibited R2 ≥0.99 over seven concentration levels with acceptable deviations.
• Workflow Efficiency: Automation reduced hands-on time, with only initial and final user interventions required.

Benefits and Practical Applications

Automation of amino acid analysis enhances laboratory throughput and reproducibility, supports quality control in bioprocessing, and enables rapid response to process deviations. The flexible 3 × 32 reagent format and standardized scripts facilitate adoption across diverse platforms.

Future Trends and Applications

Advancements may include integration with real-time monitoring systems, expanded panels for modified amino acids, and cloud-based data analytics. Further optimization of scripts on emerging liquid handling technologies will broaden applicability and streamline biopharmaceutical development pipelines.

Conclusion

A fully automated AccQ-Tag Ultra workflow provides precise, accurate, and linear quantitation of 26 amino acids in cell culture standards. The solution offers comparable performance to manual methods while significantly reducing manual labor and improving consistency across multiple platforms.

References

• Danielle Cullen et al. High Throughput Preparation of Amino Acids Using Waters Technologies AccQ-Tag Ultra Automation Kit and Cell Culture Standards. Waters Corporation, 2020.