الفهرس 
Peptide mapping of recombinant therapeutic proteins and its applications in quality assessment
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Abstract
Peptide mapping (PM) is a vital technique in biopharmaceutical industry. The fingerprint obtained helps to qualitatively confirm host stability as well as verify primary structure, purity and integrity of the target protein. Yet, in-solution digestion followed by tandem mass spectrometry is not suitable as a routine quality control test. It is time consuming and requires sophisticated, expensive instruments and highly skilled operators. In an attempt to enhance the functionality of PM and extract multi-dimensional data about various critical quality attributes, coupling of PM/UV detection to principal component analysis (PCA) is proposed. Samples of different recombinant human growth hormone (rhGH) products as well as degraded samples: deamidated and oxidized were subjected to trypsin digestion and analyzed by RP-HPLC. The entire chromatograms of test and reference samples was then used to perform PCA. Comparison of the scores of samples and investigation of the loadings plots clearly indicated the applicability of PM-PCA for: i) identity testing, ii) biosimilarity assessment and iii) stability evaluation. Hotelling{u2019}s T2 and Q statistics were employed at 95% confidence level to measure the variation and to test the conformance of each sample to the PCA model, respectively. Coupling of PM to PCA provided a novel tool to identify peptide fragments responsible for variation between the test and reference samples as well as evaluation of the extent and relative significance of this variability. Transformation of conventional PM that is largely based on subjective visual comparison into an objective statistically-guided analysis framework should provide a simple and economic tool to help both manufacturers and regulatory authorities in quality and biosimilarity assessment of biopharmaceuticals