세포 배양에서 핵심 영양분의 결정 및 통제는 단백질 생산을 극대화하고 일관된 생산 프로세스를 제공하는 데 중요한 요인입니다. Waters의 UPLC® Amino Acid Analysis Solution은 정확한 고처리량 아미노산 분석을 제공하는 턴키 솔루션입니다.
UPLC Amino Acid Analysis Solution은 단백질 특성화, 세포 배양 모니터링, 식품 및 사료의 영양 분석의 특정 요구를 충족합니다. 이 엔드-투-엔드 시스템은 사용자가 필요로 하는 정확성과 신뢰성을 제공하기 위해 개발되었습니다.
UPLC Amino Acid Analysis Solution
UPLC® Amino Acid Analysis Solution은 아미노산 분석에서 흔히 변동성을 유발하는 주요 요소를 제어합니다. 최적화된 기기와 품질 관리된 컬럼부터 표준화된 소프트웨어 프로젝트, 분석법, 보고서 형식, 퀄리피케이션에 이르기까지, 견고하고 신뢰성 있는 분석법, 즉 모든 시료의 성공적인 분석을 보장하는 종합적이고 구체적인 접근법을 제공합니다.
UPLC AAA (Amino Acid Analysis) Solution은 워터스의 업계를 선도하는 기술로, 기기, 컬럼, 소프트웨어, 퀄리피케이션, 서비스를 제공하며, 애플리케이션 중심의 뛰어난 성능에 최적화된 솔루션입니다. 과학자를 위해, 과학자에 의해 개발된 이 솔루션을 사용하면, 애플리케이션 니즈를 충족시킬 수 있는 최고의 솔루션을 선택함에 있어서 추측이나 짐작이 필요 없습니다.
Importance of Amino Acids
Analysis of amino acids is required in several areas of research, and it is a fundamental tool in product analysis. The application imposes different requirements on the analytical method because the amino acids play different roles.
With the UPLC® Amino Acid Analysis Solution, you benefit from better resolution and sensitivity, all achieved in a shorter analysis time than with conventional methodologies:
Optimized Peak Identification
AccQ•Tag™ Ultra Chemistries used in the UPLC Amino Acid Analysis Solution are a comprehensive and fully-tested set of reagents, columns, and eluents optimized for use with the ACQUITY UPLC, ACQUITY UPLC H-Class, and ACQUITY UPLC H-Class Bio Systems. With superior resolution of all amino acids, you will confidently achieve peak identification and quantitation, which in turn assures the accuracy of your results.
This chemistry is based on Waters’ widely-used and understood AccQ•Tag derivatizaton method. Primary and secondary amino acids react with a single reagent, AccQ•Fluor™, in a high-throughput batch process resulting in exceptionally stable derivatives. High resolution separations are achieved using pre-qualified AccQ•Tag Ultra UPLC columns and mobile phases. Derivatized amino acids are quantified to sub-picomole levels with single wavelength UV detection.
Qualification services available for the UPLC Amino Acid Analysis Solution meet the most stringent regulatory requirements. Installation, Operation, and Performance Qualification (IQ/OQ/PQ) protocols ensure that every element of the system is performing according to its intended use. An additional PQ, specific for amino acid analysis, is also available to verify performance and ensure accurate results.
Analysis of a Hydrolysate Standard at 10 pmol/mL
The UPLC Amino Acid Analysis Solution provides high resolution to ensure method robustness with good quantitation. The short cycle time is obtained without compromising accurate results.
Efficient, Reliable Foods and Feed Sample Analysis: Soybean Meal Hydrolysate
Two samples of hydrolyzed soybean meal were analyzed. The upper trace shows the untreated sample while the lower trace shows the results of performic acid oxidation. Note the appearance of cysteic acid and methionine sulfone in the oxidized sample.
AAA Analysis for Biopharmaceutical Applications
Accurate, precise, and robust amino acid analysis is required as fundamental information in several kinds of investigational projects as well as in routine QC and process monitoring. In biochemical structural analysis, the amino acid composition of hydrolyzed proteins is used to confirm sequence and to determine extinction coefficients. In contrast to measurements of protein bound amino acids, free amino acids are important indicators of metabolic status and genetic origin. Such analyses are required for diverse applications such as optimizing cell culture conditions in the production of biopharmaceuticals as well as process monitoring in industrial scale fermentations.
The demand of these applications led to the invention of amino acid analyzers and the continuous improvement of the technology over several decades. A turnkey, total application solution was developed. The technique based on a well-characterized derivatization protocol followed by a high resolution chromatographic separation. The amino acids are derivatized with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate (AQC) under largely aqueous conditions. The derivatives are separated using UltraPerformance Liquid Chromatography (UPLC). The enhanced resolution of UPLC ensures a robust chromatographic method that reliably gives accurate and precise measurements.
This technique has been widely adopted by biopharmaceutical laboratories since its introduction in 2007. This UPLC Amino Acid Analysis System solution is now available for Waters ACQUITY UPLC (binary) and ACQUITY UPLC H-Class and H-Class Bio (quaternary) configurations and detection protocols, with documented linearity, accuracy, and precision. The continuous enhancement of this application solution ensures the most convenient and efficient generation of reliable amino acid analyses.
AAA for Proteins, Cell Culture Media, and Feeds
The analysis of amino acids is essential in a variety of investigational projects as well as in routine QC and process monitoring. Successful analysis requires consideration of sample handling, separation and detection of the amino acids. The UPLC Amino Acid Analysis System is intended to provide accurate and precise analysis over a wide range of sample types. The amino acids are derivatized by reaction with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate and separated using reversed phase UPLC. The suitability of this method has been validated across several applications.
The amino acid composition of acid hydrolyzed pure proteins was compared to expected values from the known sequence. Amino acids as metabolic indicators were measured in growing cell culture media and various foods and beverages. The nutritional content of feeds was also determined after hydrolysis. Optimization of handling for each sample type provided more reliable results. The extreme sensitivity of the derivatization and separation techniques simplified some aspects of sample preparation while facilitating optimization of others. (See Posters under Literature tab)
Particularly for feed samples, the use of microwave hydrolysis techniques provided a practical addition to the overall analytical process. Feed samples were subjected to acid hydrolysis both with and without performic acid oxidation as well alkaline hydrolysis for a full profile of amino acid and, therefore, protein content. Sample handling throughput was much higher with this approach and was compatible with the analytical throughput of the system.
These experiments demonstrate the suitability of sample handling procedures and the amino acid analysis system
Analysis of Hydrolysate and Cell Culture Standards
The upper trace shows the analysis of a hydrolysate standard at 10 pmol/mL, and the lower trace a standard mixture of the larger number of amino acids found in cell culture media, also at 10 pmol/mL. The adaptation of the method requires only a change in dilution of the Eluent A Concentrate supplied as part of the total system solution.
Cell Culture Analysis
Direct Cell Culture Sample Analysis
Aliquots of cell culture media were taken directly from a bioreactor producing a biopharmaceutical protein. Samples were taken over a period of six days and no special sample preparation was required before the amino acid analysis shown here. The amino acid concentration trends can be plotted with growth as well as the response to feeding.