Creative Biolabs have established a platform to analyze amino acid composition of therapeutic drug candidates, strictly following the ICH Topic Q6b guideline. The overall amino acid composition of the therapeutic drug candidate is evaluated by a variety of hydrolytic and analytical procedures, and compared with the amino acid composition deduced from the DNA sequence for the desired expression product, or the natural counterpart, when necessary. In many cases, amino acid composition analysis offers structural information only for peptides and small proteins, and such data are generally less definitive for larger protein products. Quantitative amino acid analysis data can be utilized to detect protein content in many cases.
An inherent degree of heterogeneity can be produced in proteins due to the biosynthetic processes utilized by living organisms. Therefore, the manufacturing product could be a mixture of desired product and other contaminants. These contaminants may have no baneful effect on the safety and efficacy of the product. However, the manufacturer should determine the pattern of heterogeneity of the desired product and exhibit consistency with that of the lots utilized in preclinical/clinical studies. When the consistency pattern of product heterogeneity is demonstrated, determination of the bioactivity, efficacy, safety and immunogenicity of individual batches may not be necessary. Heterogeneity also occurs during manufacture and/or during storage of the drug substance or drug product. The heterogeneity of the products determine their quality, hence the level and profile of this heterogeneity should be evaluated, to maintain lot-to-lot consistency. If the variants of the desired product have characteristics comparable to those desired product in respect of bioactivity, efficacy and safety, they are defined as product related substances. When manufacturing process changes and degradation products lead to heterogeneity patterns, which differ from those utilized during pre-clinical and clinical development, the significance of these alterations should be determined.
Creative Biolabs have established a platform to determine the amino acid composition by performing hydrolysis of monoclonal antibody followed by derivation and analysis by UPLC with fluorescence detection. The obtained data can be utilized to assess amino acid molar ratio and estimate the protein concentration and extinction coefficient. Analysis data covers triplicate hydrolysis and duplicate injections of each hydrolysate (6 data points), along with UV spectrophotometry at 280nm (6 analyses in parallel). All the proteins are characterized in house in 2-3 weeks.