{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,8]],"date-time":"2026-02-08T11:14:28Z","timestamp":1770549268010,"version":"3.49.0"},"reference-count":43,"publisher":"Public Library of Science (PLoS)","issue":"8","license":[{"start":{"date-parts":[[2023,8,9]],"date-time":"2023-08-09T00:00:00Z","timestamp":1691539200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Lupin Limited"}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS ONE"],"abstract":"<jats:p>Pegfilgrastim is administered as an adjunct to chemotherapy to reduce the incidence of febrile neutropenia and associated infectious complications. Lupin\u2019s Pegfilgrastim is a proposed biosimilar to the U.S.-referenced Neulasta\u00ae. Demonstration of biosimilarity requires extensive physicochemical and functional characterization of the biosimilar, and demonstration of analytical similarity to the reference product, in addition to clinical studies. This work is a case study for demonstrating the analytical similarity of Armlupeg (Lupin\u2019s Pegfilgrastim) to Neulasta\u00ae with respect to structural and physicochemical attributes using several robust, orthogonal, and state-of-the-art techniques including high-end liquid chromatography, mass spectrometry, and spectroscopy techniques; circular dichroism; differential scanning calorimetry; nuclear magnetic resonance; analytical ultracentrifugation; and micro-flow imaging. Functional similarity was demonstrated using an <jats:italic>in vitro<\/jats:italic> cell proliferation assay to measure relative potency and surface plasmon resonance to measure receptor binding kinetics. Furthermore, comparative forced-degradation studies were performed to study the degradation of the products under stress conditions. The product attributes were ranked based on a critical quality attributes risk score according to their potential clinical impact. Based on criticality, all analyses were statistically evaluated to conclude analytical similarity. Lupin\u2019s Pegfilgrastim was comparable to Neulasta\u00ae as demonstrated via structural, functional, and purity analyses. Lupin\u2019s Pegfilgrastim complied with the quality and statistical ranges established using Neulasta\u00ae. Both products follow the same degradation pathways under stress conditions as observed in the forced-degradation studies. No new impurity or degradation product was observed in Lupin\u2019s Pegfilgrastim. These data conclusively demonstrate the analytical similarity of Lupin\u2019s Pegfilgrastim and Neulasta\u00ae.<\/jats:p>","DOI":"10.1371\/journal.pone.0289745","type":"journal-article","created":{"date-parts":[[2023,8,9]],"date-time":"2023-08-09T17:32:58Z","timestamp":1691602378000},"page":"e0289745","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":5,"title":["Analytical sameness methodology for the evaluation of structural, physicochemical, and biological characteristics of Armlupeg: A pegfilgrastim biosimilar case study"],"prefix":"10.1371","volume":"18","author":[{"given":"Arati","family":"Deshmukh","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rishank","family":"Goyal","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kalyana","family":"Sundaram","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kaustubh","family":"Dange","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tejshri","family":"Lakhote","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sanjay","family":"Niranjan","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jennifer","family":"Bharucha","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ashok","family":"Mishra","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3141-0355","authenticated-orcid":true,"given":"Bhavesh","family":"Vats","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sanjay","family":"Tiwari","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"340","published-online":{"date-parts":[[2023,8,9]]},"reference":[{"key":"pone.0289745.ref001","doi-asserted-by":"crossref","first-page":"1235","DOI":"10.2174\/1381612043452613","article-title":"The Design and Development of Pegfilgrastim (PEG-rmetHuG-CSF, Neulasta&#174;).","volume":"10","author":"G. 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Lupin Limited; 2020."},{"key":"pone.0289745.ref006","doi-asserted-by":"crossref","first-page":"410","DOI":"10.3390\/biom9090410","article-title":"The Breakthrough of Biosimilars: A Twist in the Narrative of Biological Therapy.","volume":"9","author":"ER Kabir","year":"2019","journal-title":"Biomolecules."},{"key":"pone.0289745.ref007","doi-asserted-by":"crossref","first-page":"iv14","DOI":"10.1093\/rheumatology\/kex278","article-title":"The process defines the product: what really matters in biosimilar design and production?","volume":"56","author":"AG Vulto","year":"2017","journal-title":"Rheumatology."},{"key":"pone.0289745.ref008","first-page":"541","article-title":"Pegfilgrastim Biosimilars: Where Are We Now?","volume":"12","author":"C Selby","year":"2021","journal-title":"J Adv Pract Oncol."},{"key":"pone.0289745.ref009","article-title":"Comparison of Efficacy and Safety of Lupin\u2019s Pegfilgrastim with Neulastim\u00ae as an Adjunct to Chemotherapy in Patients with Non Myeloid Malignancies: A Randomised Phase III Clinical Study","volume":"16","author":"M Jain","year":"2022","journal-title":"JOURNAL OF CLINICAL AND DIAGNOSTIC RESEARCH"},{"key":"pone.0289745.ref010","doi-asserted-by":"crossref","first-page":"1910","DOI":"10.1016\/j.drudis.2020.09.006","article-title":"Streamlined approval of biosimilars: moving on from the confirmatory efficacy trial.","volume":"25","author":"M-C Bielsky","year":"2020","journal-title":"Drug Discov Today"},{"key":"pone.0289745.ref011","doi-asserted-by":"crossref","first-page":"265","DOI":"10.1080\/14740338.2021.1876024","article-title":"Considerations related to comparative clinical studies for biosimilars.","volume":"20","author":"AS Rathore","year":"2021","journal-title":"Expert Opin Drug Saf"},{"key":"pone.0289745.ref012","doi-asserted-by":"crossref","first-page":"359","DOI":"10.1007\/s40259-022-00533-x","article-title":"Regulatory Evaluation of Biosimilars: Refinement of Principles Based on the Scientific Evidence and Clinical Experience.","volume":"36","author":"P Kurki","year":"2022","journal-title":"BioDrugs."},{"key":"pone.0289745.ref013","doi-asserted-by":"crossref","first-page":"199","DOI":"10.1016\/j.yrtph.2015.12.013","article-title":"Biosimilars: A consideration of the regulations in the United States and European union","volume":"76","author":"J. Daller","year":"2016","journal-title":"Regulatory Toxicology and Pharmacology"},{"key":"pone.0289745.ref014","unstructured":"Development of Therapeutic Protein Biosimilars: Comparative Analytical Assessment and Other Quality-Related Considerations Guidance for Industry DRAFT GUIDANCE. 2019. 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Deutzmann","year":"2004","journal-title":"Methods Mol Med"},{"key":"pone.0289745.ref022","doi-asserted-by":"crossref","first-page":"349","DOI":"10.2174\/1389203003381315","article-title":"The Use of Circular Dichroism in the Investigation of Protein Structure and Function.","volume":"1","author":"S Kelly","year":"2000","journal-title":"Curr Protein Pept Sci."},{"key":"pone.0289745.ref023","doi-asserted-by":"crossref","first-page":"1655","DOI":"10.1002\/elps.202100045","article-title":"A simple, rapid, and robust \u201con\u2010the\u2010go\u201d identity testing of biotherapeutics using FTIR spectroscopy.","volume":"42","author":"MA Afzal","year":"2021","journal-title":"Electrophoresis"},{"key":"pone.0289745.ref024","doi-asserted-by":"crossref","first-page":"432","DOI":"10.1021\/ar700188n","article-title":"Amide I Two-Dimensional Infrared Spectroscopy of Proteins","volume":"41","author":"Z Ganim","year":"2008","journal-title":"Acc Chem Res"},{"key":"pone.0289745.ref025","doi-asserted-by":"crossref","first-page":"22518","DOI":"10.3390\/ijms151222518","article-title":"Intrinsic Tryptophan Fluorescence in the Detection and Analysis of Proteins: A Focus on F\u00f6rster Resonance Energy Transfer Techniques.","volume":"15","author":"A Ghisaidoobe","year":"2014","journal-title":"Int J Mol Sci"},{"key":"pone.0289745.ref026","doi-asserted-by":"crossref","first-page":"163","DOI":"10.1007\/s12551-016-0197-7","article-title":"UV\u2013Vis spectroscopy of tyrosine side-groups in studies of protein structure. Part 2: selected applications.","volume":"8","author":"JM Antosiewicz","year":"2016","journal-title":"Biophys Rev."},{"key":"pone.0289745.ref027","doi-asserted-by":"crossref","first-page":"2467","DOI":"10.1007\/s00216-017-0772-1","article-title":"Recent mass spectrometry-based techniques and considerations for disulfide bond characterization in proteins","volume":"410","author":"JC Lakbub","year":"2018","journal-title":"Anal Bioanal Chem"},{"key":"pone.0289745.ref028","doi-asserted-by":"crossref","DOI":"10.1016\/bs.mie.2015.08.029","article-title":"Orthogonal Methods for Characterizing the Unfolding of Therapeutic Monoclonal Antibodies","author":"DB Temel","year":"2016","journal-title":"Methods Enzymol"},{"key":"pone.0289745.ref029","doi-asserted-by":"crossref","first-page":"6366","DOI":"10.1021\/acs.analchem.9b05385","article-title":"Comparative Analysis of One-Dimensional Protein Fingerprint by Line Shape Enhancement and Two-Dimensional 1 H, 13 C Methyl NMR Methods for Characterization of the Higher Order Structure of IgG1 Monoclonal Antibodies","volume":"92","author":"KW Elliott","year":"2020","journal-title":"Anal Chem"},{"key":"pone.0289745.ref030","doi-asserted-by":"crossref","first-page":"4251","DOI":"10.3390\/molecules26144251","article-title":"NMR Spectroscopy for Protein Higher Order Structure Similarity Assessment in Formulated Drug Products.","volume":"26","author":"D Wang","year":"2021","journal-title":"Molecules"},{"key":"pone.0289745.ref031","doi-asserted-by":"crossref","first-page":"6559","DOI":"10.1007\/s00216-013-7469-x","article-title":"Analytical techniques and bioactivity assays to compare the structure and function of filgrastim (granulocyte-colony stimulating factor) therapeutics from different manufacturers.","volume":"406","author":"MJ Levy","year":"2014","journal-title":"Anal Bioanal Chem"},{"key":"pone.0289745.ref032","doi-asserted-by":"crossref","first-page":"114804","DOI":"10.1016\/j.ab.2022.114804","article-title":"Utility of SPR technology in biotherapeutic development: Qualification for intended use","volume":"654","author":"W Wang","year":"2022","journal-title":"Anal Biochem"},{"key":"pone.0289745.ref033","doi-asserted-by":"crossref","first-page":"297","DOI":"10.1007\/s40259-020-00422-1","article-title":"The Path Towards a Tailored Clinical Biosimilar Development.","volume":"34","author":"M Schiestl","year":"2020","journal-title":"BioDrugs"},{"key":"pone.0289745.ref034","doi-asserted-by":"crossref","first-page":"895","DOI":"10.1002\/jps.22812","article-title":"Forced Degradation of Therapeutic Proteins","volume":"101","author":"A Hawe","year":"2012","journal-title":"J Pharm Sci"},{"key":"pone.0289745.ref035","doi-asserted-by":"crossref","first-page":"291","DOI":"10.1016\/j.biologicals.2016.06.005","article-title":"Determination of critical quality attributes for monoclonal antibodies using quality by design principles.","volume":"44","author":"N Alt","year":"2016","journal-title":"Biologicals"},{"key":"pone.0289745.ref036","doi-asserted-by":"crossref","first-page":"670","DOI":"10.1208\/s12248-016-9882-5","article-title":"Analytical Similarity Assessment in Biosimilar Studies.","volume":"18","author":"S-C Chow","year":"2016","journal-title":"AAPS J."},{"key":"pone.0289745.ref037","doi-asserted-by":"crossref","first-page":"28","DOI":"10.1016\/j.biologicals.2017.06.001","article-title":"A demonstration of analytical similarity comparing a proposed biosimilar pegfilgrastim and reference pegfilgrastim.","volume":"48","author":"S Brokx","year":"2017","journal-title":"Biologicals"},{"key":"pone.0289745.ref038","doi-asserted-by":"crossref","first-page":"e0212622","DOI":"10.1371\/journal.pone.0212622","article-title":"Structural similarity, characterization of Poly Ethylene Glycol linkage and identification of product related variants in biosimilar pegfilgrastim.","volume":"14","author":"R Shekhawat","year":"2019","journal-title":"PLoS One"},{"key":"pone.0289745.ref039","unstructured":"Biosimilar Product Information. In: U.S. Food & Drug Administration. 19 Dec 2022."},{"key":"pone.0289745.ref040","doi-asserted-by":"crossref","first-page":"105501","DOI":"10.1016\/j.ejps.2020.105501","article-title":"Reporting of quality attributes in scientific publications presenting biosimilarity assessments of (intended) biosimilars: a systematic literature review.","volume":"154","author":"AM Alsamil","year":"2020","journal-title":"European Journal of Pharmaceutical Sciences"},{"key":"pone.0289745.ref041","doi-asserted-by":"crossref","first-page":"321","DOI":"10.1007\/s40259-016-0184-3","article-title":"Assessing Analytical Similarity of Proposed Amgen Biosimilar ABP 501 to Adalimumab.","volume":"30","author":"J Liu","year":"2016","journal-title":"BioDrugs."},{"key":"pone.0289745.ref042","doi-asserted-by":"crossref","first-page":"114","DOI":"10.1007\/s11095-020-02816-w","article-title":"Analytical and Functional Similarity Assessment of ABP 710, a Biosimilar to Infliximab Reference Product","volume":"37","author":"R Saleem","year":"2020","journal-title":"Pharm Res"},{"key":"pone.0289745.ref043","doi-asserted-by":"crossref","first-page":"62","DOI":"10.3390\/bioengineering6030062","article-title":"Forced Degradation Testing as Complementary Tool for Biosimilarity Assessment.","volume":"6","author":"YFK Dyck","year":"2019","journal-title":"Bioengineering."}],"container-title":["PLOS ONE"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0289745","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,8,9]],"date-time":"2023-08-09T17:34:15Z","timestamp":1691602455000},"score":1,"resource":{"primary":{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0289745"}},"subtitle":[],"editor":[{"given":"Krishna M. 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