Rapid Identification and Characterization of Formulated Protein Products by Raman Spectroscopy Coupled with Discriminant Analysis.

UNLABELLED: The rapid identification of protein drug products for packaging and receiving can significantly reduce disposition cycle time, and thereby improve the efficiency and productivity of the supply chain to better meet the needs of patients. In this feasibility study, we demonstrate a novel methodology that combines Raman spectroscopy with discriminant analysis that can be used for rapid identification or verification of finished products. With this methodology, Raman spectra of formulated therapeutic proteins were collected non-invasively with the samples either in a quartz cuvette or in the original glass vials, and analyzed without subtraction of buffer or placebo solutions. The algorithm used for the discriminant analysis was Mahalanobis distance by principal component analysis with residuals. In addition to product identification, the methodology has the potential to be used for characterizing formulated proteins when exposed to external stresses based on the changes of Mahalanobis distances. LAY ABSTRACT: The rapid identification of protein drug products for packaging and receiving can significantly reduce disposition cycle time, and thereby improve the efficiency and productivity of the supply chain. In this study, we demonstrate a novel methodology that combines Raman spectroscopy with discriminant analysis to rapidly identify formulated proteins non-invasively.

Routine screening for the presence of adulteration in raw materials using automated nuclear magnetic resonance spectroscopy.

In an effort to increase the security of the supply chain for raw materials used in the manufacture of human therapeutics, a routine screen to detect the presence of adulteration using fully automated nuclear magnetic resonance spectroscopy has been developed and qualified for use in quality control laboratories. The method involves the collection of one-dimensional (1)H and (13)C spectra, which are subsequently processed to identify and quantitate raw material constituents by comparison to a spectral database. The resulting method is an easy-to-use limit test that can automatically determine the integrity of incoming raw materials. The method is intended to be used in good manufacturing practice production facilities and is suitable for excipients and aqueous soluble raw materials used in biopharmaceutical processes. LAY ABSTRACT: In an effort to increase the security of the supply chain for raw materials used in the manufacture of human therapeutics, a routine screen to detect the presence of adulteration using fully automated nuclear magnetic resonance (NMR) spectroscopy has been developed and qualified for use in quality control laboratories. The method involves the collection of NMR spectra, which are subsequently processed to identify and quantitate raw material constituents by comparison to a spectral database. The resulting method is an easy-to-use limit test that can automatically determine the integrity of incoming raw materials. The method is intended to be used in good manufacturing practice production facilities and is suitable for excipients and aqueous soluble raw materials used in biopharmaceutical processes.

Biochemical assessment of erythropoietin products from Asia versus US Epoetin alfa manufactured by Amgen.

We compared the physical and chemical properties of purported copies of recombinant human erythropoietin (rHuEPO) purchased from Korea, China, and India with the innovator product, Epoetin alfa, manufactured by Amgen Inc. The products were characterized for similarity in the types of glycoforms present, the relative degree of unfolding, in vitro potency, presence of covalent aggregates, and presence of cleavage products using established analytical methods. All products were different from Epoetin alfa (Epogen). The purported copies of rHuEPO from Korea, India, and China contained more glycoforms and other impurities. The in vitro relative potency varied for each product when based on the labeled concentration, while the concentration based on ELISA analysis brought the relative potency, for most products closer to 100%. These data emphasize potential biochemical discrepancies resulting from different cell lines and manufacturing processes. Concentrations varied within products and did not always match the information provided on the product label. As it is not possible to reliably correlate such biochemical discrepancies to clinical consequences, or the lack thereof, these data support the need for extensive preclinical testing and clinical testing of all investigational products as not all safety and efficacy aspects can be assessed during preclinical evaluation.