Amino Acid Analysis Service
Amino Acid Analysis (AAA) is essential for understanding the protein composition of your biopharmaceutical product and showing control over your process through demonstrating manufacturing consistency across batches.
At BioPharmaSpec, we specialize in AAA and extinction coefficient determination, delivering precise and dependable results tailored to your requirements and regulatory standards.
- Total Protein Content
- Extinction coefficient estimation
- Extinction coefficient determination
- Free amino acid quantitation
- D-amino acid detection
- Customized method development (unusual/non-proteinogenic amino acids)
The BioPharmaSpec Advantage
- Industry-Leading Expertise: Our team comprises pioneering industry experts with extensive experience in sample handling and preparation for Amino Acid Analysis.
- Cutting-Edge Technology: Utilizing state-of-the-art instrumentation and methods, BioPharmaSpec ensures the highest quality analysis for your biopharmaceutical. Our technology allows for the efficient quantification of amino acids using UPLC with fluorescence detection.
- Proven Track Record: With over a decade of experience, we've consistently delivered cutting-edge analytical solutions to biotech enterprises, Contract Manufacturing Organizations (CMOs) and pharmaceutical companies throughout the World.
- Customized Service: We provide professional assistance throughout your project, offering method development to suit your specific requirements. Our ability to customize exclusive solutions based on your specific protein ensures optimal outcomes.
Our Amino Acid Analysis Methodology
To analyze the amino acid composition of a protein, our process begins with the breakdown of the polypeptide into its constituent amino acids. Here's how we perform Amino Acid Analysis:
- Acid Hydrolysis: We initiate the process with a 6N HCL acid hydrolysis.
- Pre-column Derivatization Chemistry: Next, we employ automated pre-column derivatization chemistry using OPA (ortho-phthalaldehyde) for primary amino acids and FMOC (9-fluorenymethyl chloroformate) for secondary amino acids.
- Separation and Detection: These amino acids are then separated using UPLC (Ultra-Performance Liquid Chromatography) with fluorescence detection.
- Quantification: Specific amino acids in a protein sample are accurately quantified by comparing them to data obtained from a standard mixture containing known amounts.
The ICH Q6B guidelines require that amino acid compositional analysis be performed as part of a complete structural characterization package. The guidelines state that “Quantitative Amino Acid Analysis can also be used to determine protein content in many cases.”
Estimation of Extinction Coefficient Through Amino Acid Analysis
At BioPharmaSpec, we utilize the OPA/FMOC pre-column derivatization method to evaluate amino acid molar ratios and estimate protein concentration and extinction coefficient. Before analysis, the Optical Density (OD) of a sample aliquot is measured at 280 nm against a suitable buffer blank.
When utilizing AAA data, several factors must be considered. For instance, individual amino acids vary in susceptibility to acid hydrolysis. Longer hydrolyses yield more accurate results for hydrophobic amino acids like Valine, Isoleucine, and Phenylalanine. Asparagine and Glutamine convert directly into corresponding acids, affecting direct quantification. Losses of amino acids like Serine, Threonine, Methionine, and Tyrosine can be expected through hydrolytic breakdown. Cysteine determination is challenging, due to oxidation and Tryptophan is completely destroyed during normal hydrolysis.
For optimal AAA data, it's crucial to work with experienced scientists familiar with this methodology.
Streamlined Analytical Process
Our approach to amino acid composition analysis is designed for efficiency, transparency and excellence at every stage. From initial consultation to ongoing support, we prioritize the success of your project, ensuring results that exceed expectations.
What is the extinction coefficient of my product?
Ready to optimize your biopharmaceutical development process? Let's start with a conversation. Contact our scientists to discuss your project requirements and receive a quote for performing AAA or estimating/determining the extinction coefficient of your biopharmaceutical.