Analytical strategies for assessing comparability of biosimilars

Biosimilar products are required by regulatory authorities to have appropriate and comparable quality, safety and efficacy with a reference biologic product. The analytical techniques that are available to manufacturers of biosimilar medicines are well advanced and present a range of options to characterize the products and compare them with the relevant marketed reference product. This variety of analytical methods can be used to investigate the attributes of a candidate biosimilar and a reference biologic, and ensures that orthogonal methods can be deployed to characterize a single attribute, facilitating rigorous structural characterization and physicochemical evaluation. These analytical methodologies not only enable evidence for bio-comparability to be amassed but also enable a developer of biosimilars to incorporate risk mitigation into the development strategy. This review discusses the analytical techniques available to support comparability testing, the handling of reference material and the strategic aspects to commercializing biosimilar or follow-on biological medicinal products.

[1]  A. Shevchenko,et al.  Femtomole sequencing of proteins from polyacrylamide gels by nano-electrospray mass spectrometry , 1996, Nature.

[2]  W. Caughey,et al.  Protein secondary structures in water from second-derivative amide I infrared spectra. , 1990, Biochemistry.

[3]  M. V. Regenmortel,et al.  Mapping Epitope Structure and Activity: From One-Dimensional Prediction to Four-Dimensional Description of Antigenic Specificity , 1996 .

[4]  S. Lee-Huang.  Cloning and expression of human erythropoietin cDNA in Escherichia coli. , 1984, Proceedings of the National Academy of Sciences of the United States of America.

[5]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.

[6]  D. Chang,et al.  Mapping of the active site of recombinant human erythropoietin. , 1997, Blood.

[7]  M. Barkley,et al.  Toward understanding tryptophan fluorescence in proteins. , 1998, Biochemistry.

[8]  Yow-Ming C Wang,et al.  Development of Biosimilars—Pharmacokinetic and Pharmacodynamic Considerations , 2009, Journal of biopharmaceutical statistics.

[9]  C. Roberts.  Kinetics of Irreversible Protein Aggregation: Analysis of Extended Lumry−Eyring Models and Implications for Predicting Protein Shelf Life , 2003 .

[10]  D G Myszka,et al.  Advances in surface plasmon resonance biosensor analysis. , 2000, Current opinion in biotechnology.

[11]  A. Vermeer,et al.  The thermal stability of immunoglobulin: unfolding and aggregation of a multi-domain protein. , 2000, Biophysical journal.

[12]  T. Arakawa,et al.  Protein isolated from biopharmaceutical formulations cannot be used for comparative studies: Follow-up to "a case study using Epoetin Alfa from Epogen and EPREX". , 2007, Journal of pharmaceutical sciences.

[13]  S. Elliott,et al.  Control of rHuEPO biological activity: the role of carbohydrate. , 2004, Experimental hematology.

[14]  B. Wallace,et al.  Protein secondary structure analyses from circular dichroism spectroscopy: methods and reference databases. , 2008, Biopolymers.

[15]  S. Demarest,et al.  A broad range of Fab stabilities within a host of therapeutic IgGs. , 2007, Biochemical and biophysical research communications.

[16]  Gopi Shankar,et al.  A risk-based bioanalytical strategy for the assessment of antibody immune responses against biological drugs , 2007, Nature Biotechnology.

[17]  D. Myszka,et al.  Kinetic analysis of macromolecular interactions using surface plasmon resonance biosensors. , 1997, Current opinion in biotechnology.

[18]  Jill Crouse-Zeineddini,et al.  Biochemical assessment of erythropoietin products from Asia versus US Epoetin alfa manufactured by Amgen. , 2009, Journal of pharmaceutical sciences.

[19]  R. Adami,et al.  Accelerated aging: prediction of chemical stability of pharmaceuticals. , 2005, International journal of pharmaceutics.

[20]  Brian M. Murphy,et al.  Stability of Protein Pharmaceuticals: An Update , 2010, Pharmaceutical Research.

[21]  J. Fitter,et al.  How aggregation and conformational scrambling of unfolded states govern fluorescence emission spectra. , 2006, Biophysical journal.

[22]  E. Lewis,et al.  Isothermal titration calorimetry: experimental design, data analysis, and probing macromolecule/ligand binding and kinetic interactions. , 2008, Methods in cell biology.

[23]  Hongwei Xie,et al.  Rapid comparison of a candidate biosimilar to an innovator monoclonal antibody with advanced liquid chromatography and mass spectrometry technologies , 2010, mAbs.

[24]  E. Moors,et al.  Clinical comparability and European biosimilar regulations , 2010, Nature Biotechnology.

[25]  H. Susi,et al.  Examination of the secondary structure of proteins by deconvolved FTIR spectra , 1986, Biopolymers.

[26]  S. Shire,et al.  A critical review of analytical ultracentrifugation and field flow fractionation methods for measuring protein aggregation , 2006, The AAPS Journal.

[27]  W. Hancock,et al.  Mass spectrometric analysis of innovator, counterfeit, and follow‐on recombinant human growth hormone , 2009, Biotechnology progress.

[28]  V. Hlady,et al.  Methods for studying protein adsorption. , 1999, Methods in enzymology.

[29]  L. Medved,et al.  Domain structure and domain-domain interactions of recombinant tissue plasminogen activator. , 1991, The Journal of biological chemistry.

[30]  W. Hancock,et al.  Mass spectrometric determination of disulfide linkages in recombinant therapeutic proteins using online LC-MS with electron-transfer dissociation. , 2009, Analytical chemistry.

[31]  S. Tamilvanan,et al.  Clinical concerns of immunogenicity produced at cellular levels by biopharmaceuticals following their parenteral administration into human body , 2010, Journal of drug targeting.

[32]  A. Panitch,et al.  Identification and sequence composition characterization of chondroitin sulfate-binding peptides through peptide array screening. , 2010, Biochemistry.

[33]  Allen R Nissenson,et al.  Pure red-cell aplasia and epoetin therapy. , 2004, The New England journal of medicine.

[34]  D. Chang,et al.  Fine-structure epitope mapping of antierythropoietin monoclonal antibodies reveals a model of recombinant human erythropoietin structure. , 1996, Blood.

[35]  K. Aoki,et al.  Biophysical comparability of the same protein from different manufacturers: a case study using Epoetin alfa from Epogen and Eprex. , 2006, Journal of pharmaceutical sciences.

[36]  E. Waters,et al.  Thermal stability of thaumatin-like protein, chitinase, and invertase isolated from Sauvignon blanc and Semillon juice and their role in haze formation in wine. , 2010, Journal of agricultural and food chemistry.

[37]  W. Hancock,et al.  Identification of the unpaired cysteine status and complete mapping of the 17 disulfides of recombinant tissue plasminogen activator using LC-MS with electron transfer dissociation/collision induced dissociation. , 2010, Analytical Chemistry.

[38]  Sandeep Nema,et al.  Protein Structural Conformation and Not Second Virial Coefficient Relates to Long-Term Irreversible Aggregation of a Monoclonal Antibody and Ovalbumin in Solution , 2006, Pharmaceutical Research.

[39]  C. Djerassi,et al.  Isoelectric fractionation, analysis, and characterization of ampholytes in natural pH gradients. IV. Further studies on the resolving power in connection with separation of myoglobins. , 1966, Acta chemica Scandinavica.

[40]  P. Seeburg,et al.  Direct expression in Escherichia coli of a DNA sequence coding for human growth hormone , 1979, Nature.

[41]  A. Rozhkova.  Quantitative analysis of monoclonal antibodies by cation-exchange chromatofocusing. , 2009, Journal of chromatography. A.

[42]  C. Dempsey.  Hydrogen exchange in peptides and proteins using NMR spectroscopy , 2001 .

[43]  William F Weiss,et al.  Principles, approaches, and challenges for predicting protein aggregation rates and shelf life. , 2009, Journal of pharmaceutical sciences.

[44]  A. Middelberg,et al.  Intrinsic fluorescence as an analytical probe of virus-like particle assembly and maturation. , 2008, Biochemical and biophysical research communications.

[45]  D. Kalonia,et al.  Temperature- and pH-Induced Multiple Partially Unfolded States of Recombinant Human Interferon-α2a: Possible Implications in Protein Stability , 2003, Pharmaceutical Research.

[46]  I. Jelesarov,et al.  Survey of the year 2008: applications of isothermal titration calorimetry , 2010, Journal of molecular recognition : JMR.

[47]  W. Hancock,et al.  Characterization of the glycosylation occupancy and the active site in the follow-on protein therapeutic: TNK-tissue plasminogen activator. , 2010, Analytical chemistry.

[48]  Y. Reshetnyak,et al.  Decomposition of protein tryptophan fluorescence spectra into log-normal components. II. The statistical proof of discreteness of tryptophan classes in proteins. , 2001, Biophysical journal.

[49]  T. Harvey,et al.  NMR structure of human erythropoietin and a comparison with its receptor bound conformation , 1998, Nature Structural Biology.

[50]  John P. Gabrielson,et al.  Quantitation of aggregate levels in a recombinant humanized monoclonal antibody formulation by size-exclusion chromatography, asymmetrical flow field flow fractionation, and sedimentation velocity. , 2007, Journal of pharmaceutical sciences.

[51]  Gary Walsh,et al.  Biopharmaceutical benchmarks 2010 , 2010, Nature Biotechnology.

[52]  N. Casadevall,et al.  The increased incidence of pure red cell aplasia with an Eprex formulation in uncoated rubber stopper syringes. , 2005, Kidney international.

[53]  Gerd R Kleemann,et al.  Optimization of a reversed-phase high-performance liquid chromatography/mass spectrometry method for characterizing recombinant antibody heterogeneity and stability. , 2006, Journal of chromatography. A.

[54]  G. Stamper,et al.  Accelerated Stability Testing of Proteins and Peptides: pH-Stability Profile of Insulinotropin Using Traditional Arrhenius and Non-Linear Fitting Analysis , 1995 .

[55]  J. Philo.  Is any measurement method optimal for all aggregate sizes and types? , 2006, The AAPS Journal.

[56]  J. Greenberger,et al.  Demonstration of permanent factor-dependent multipotential (erythroid/neutrophil/basophil) hematopoietic progenitor cell lines. , 1983, Proceedings of the National Academy of Sciences of the United States of America.