3Major B&B Themes and Initiatives for 2010-2015 Modernization of MonographsOutreach to Industry for New MonographsProcedural Chapters and Characterization/Measurement of Ancillary and Process MaterialsGrowing the portfolio to support all biologicsModernization of analytical approachesBiological Potency Tests:Implementation of new USP ChaptersAssay transitions: animal to cell, cell to binding, activity to amount of substance – questions of equivalence and unitsProduct Class Chapters:Monoclonal antibodiesGlycoconjugate vaccines
4Vertical Standards – Monographs Role and Use:Clearly define identity, strength and purity, as well as other important quality attributes at the product levelAllow independent testing and verification based on a public standardConsiderations for Standard Development:Complexity of specifications and system suitability criteriaBiological potency assignments and unit maintenanceAcross manufacturersInternationallyProduct- specific vs. common product class requirementsFor well-characterized and legacy products: inclusion and bridging to new analytical technology
5Official USP Biologics Monographs by Product Class Number of monographspeptide47enzyme12complex extract11carbohydrateglycosaminoglycan9other5Tissue product6IgG/serumBlood component/proteinVaccine3
6Current Biologics in the US Market From Kozlowski et al., NEJM 265;5, 2011
10Horizontal Standards - Procedural Benefits:Access to validated procedures and procedure performance criteria early in developmentSolid anchor point for product characterizationFacilitate comparability during all development stagesDirectly apply ICH guidance, including:Q5EQ6BEstablish consistency in analytical expectations
11From General to Specific - Procedures <1084>Glycoprotein andGlycan Analysis – Introduction and Choice of Analysis MethodsGuidance & Information<212>Oligosaccharide Analysis<210>Monosaccharide AnalysisProcedures & Performance CriteriaOligosaccharide mixturesfromHuman α acid GlycoproteinFetuinRNAse BHuman IgGMonosaccharide Mixes 1–4Procedural (Horizontal) Standards for System Suitability & ValidationSingle Monosaccharides
14Test Chapters – Current Major Initiatives Impurities<30> Residual DNAPhysicochemical Tests<212> Oligosaccharide Analysis<210> Monosaccharide Analysis<121.1> Insulin Physicochemical Analysis<209> Low Molecular Weight Heparin Molecular Weight Determinations<XXX> CollagenPotency Assays and Content Measurement<57> Protein Determination Procedures<123> Glucagon Bioidentity Tests<124> Epoetin Bioassays<126> Somatropin Bioassays<208> Anti-Factor IIa and Xa Assays for Unfractionated and Low Molecular Weight Heparins
15Vaccine Chapters Possible PC Chapters <1235> Vaccines for Human Use (completed)<1238> Bacterial Vaccines (completed)<1239> Viral Vaccines (in progress)<XXXX> Toxins/Toxoids<XXXX> Live Attenuated<1234> Polysaccharide and GlycoconjugatePossible PCChapters<XXXX> Killed Viruses<XXXX> Live Attenuated<XXXX> Subunit Vaccines<XXXX> Other SubunitSub-<1000> Analytical Chapters for Key Quality Attributes and RS
16Will be accompanied by USP MAb System Suitability RS <129> Analytical Procedures for Recombinant Therapeutic Monoclonal AntibodiesWill contain a collection of validated compendial procedures with established system suitability criteria for therapeutic MAbsWill be accompanied by USP MAb System Suitability RSWill not contain product or class specific acceptance criteriaWill be supported by a suite of >1000 Information Chapters that discuss quality attributes, manufacturing and quality control aspects for MAbs
17Horizontal Standards– Ancillary & Process Materials Benefits:Quality specifications and standards for complex ancillary and process materialsGive access to process ingredients of consistent qualityControl process variabilityFacilitate testing of process intermediates and bulksChallenges in Standard Development:Fast evolution of process materials:Serum-free, proprietary custom mediaEngineered 2nd and 3rd generation materials, e.g. Protein ADefining key quality attributes for complex materials, e.g. Fetal Bovine Serum (FBS)
18Raw/Ancillary Materials Raw materials may or may not remain in the final therapeutic product as active substances or excipientsAncillary materials are a subset of raw materialsAncillary products may exert an effect on a therapeutic substance(e.g. a cytokine may activate a population of cells), but they are not intended to be in final formulationConcerns related to raw materials qualification have been amplified by recent materials supply issues:Glycerin, Heparin, MelamineAnimal derived material (serum) used in manufacturing of some biologicsSome components are more critical than others! Risk assessment strategies are required to ensure qualityA critical material will come in contact with cells with a potential to alter either the growth characteristics of the cells or the ability of the cell culture to meet lot release specifications.
19Ancillary Materials Standards: USP Approach General Information Chapter (Guidance)<1043> Ancillary Materials for Cell-, Gene-, and Tissue-Engineered ProductsSpecific AM Chapters<90> FBS Quality Attributes<92> Cytokines and Growth Factors Quality AttributesAncillary Material Requirements for Specific AMsReference Standards:FBSInterleukin-4FGF-2TransferrinG-CSFAncillary Material Reference Standards
20Risk-based Classification of Ancillary Materials Tier 1 – Low-Risk, Highly Qualified Materials with Intended Use as Therapeutic Drug or Biologic, Medical Device, or Implantable MaterialTier 2 – Low-Risk, Well Characterized Materials with Intended Use as AMs, Produced in Compliance with GMPsTier 3 – Moderate-Risk Materials Not Intended for Use as AMs (frequently produced for in vitro diagnostic use or reagent grade materials)Tier 4 – High-Risk Materials, Materials not Produced in Compliance with cGMPs and materials not intended to be used in cell manufacturing
21Risk-based Qualification – USP <1043> Elements of Qualification and/or Risk Reduction ActivitiesTier 1Tier 2Tier 3Tier 4Master File cross referenceXCertificate of analysislot-to-lot effect on process performanceRemoval from finished productStability as stored and used in specific processConfirm Certificate Analysis TestVendor AuditUpgrade Manufacturing to GMP levelDevelop internal specificationsLot to lot comparability may be neededTesting for adventitious agents may be neededTraceability to country of origin, safety from animal diseasesAdventitious agent testing for animal source-relevant viruses
22Recombinant Human Interleukin 4 -USP Interleukin-4 Standard RequirementSpecific Activity: Not less than 0.5 x 107 USP Units /mg of total proteinLabeled potency of RS will be based on bioactivity using TF-1 cell linePurity: 98% (SDS-PAGE and silver stain)Identity: N-term protein sequencing (10 residues) and Western BlotAssociated Reference StandardLyophilized powderCalibrated against International Standard (WHO)How is the standard used?Preparation of IL-4 for which the activity was determined by calibration against the USP standard, will provide consistency in the manufacturing of the cell therapy product, by way of using the right amount of material.
23USP General Chapter <90> Fetal Bovine Serum – Quality Attributes and Functionality TestsOfficial uses of the USP FBS Reference StandardIdentification – Radial ImmunodiffusionFBS Functionality Tests – Growth Promotion CurveFive cell lines are recommended for useFunctionality tests are performed on 3 cell linesTwo from the recommended cell line listThird is cell line relevant to the user’s application
24Fetal Bovine Serum (FBS)- USP FBS Standard RequirementsOsmolality: mOsm/KgTotal Protein: mg/mLpH:Endotoxin: Not more than 10 units/mLHemoglobin level: Not more than 30 mg/dLIdentification: Radial Immunodiffusion (RID): species ID, IgG levelsFunctionality Assays (Growth Curve and Clonal Assay)Associated Reference Standard (RS), under developmentLiquid frozen, 10 mLCollaborative study to include several laboratories to test:Identification (FBS sample positive for bovine IgG and content is < 500 mg/L)Growth curve (doubling time in test sample is not less than 90% compared to RS)
25How the FBS Standard is Used: Growth Curve Challenges: Cell Line, Cell Density, Cell Counting, Days in CultureThree cell densities, determine viable cell counts on days 0,1,2,3,4, and 7. Select thecell density that exhibit a growth curve with3 phases: Lag, Log, Stationary; and linearover 3 time points or moreUse the selected cell density to assess thetest FBS side by side with the referencestandard FBSDoubling time is estimated using a growthcurve that is linear over three or more timepoints.Acceptance Criteria:R2≥ 0.98Doubling time of test sample should be notless than 90% of doubling time of RS
26Future Directions for USP Ancillary Materials Standards Growth Factors and Cytokines – additions to <92>.Next revision adds FGFStandards for enzymes as ancillary materialsTrypsin, others?Second and third generation identification tests for complex, naturally derived materials:Application of modern immunology and proteomics approaches?
27Thank you!As you can see from the world map on the outside of USP headquarters building, it is a global pharmacopeia based in Rockville, Maryland. No borders.