INTERTEK PHARMACEUTICAL SERVICES BIOPHARMACEUTICAL SERVICES

Expert Analytical & Bioanalytical Services INTERTEK BIOANALYSIS SERVICES

Our Biopharmaceutical Expertise

To meet your development milestones and comply with regulatory requirements, you will need high quality bioanalytical and analytical data to aid informed decision-making and identify sources of risk. Safety assessment is key, as slight changes in the structure, physicochemical properties, stability and impurity profile of a biologic can provoke an adverse immune response.

Our thought-leaders have over 25 years Our biopharmaceutical services Our experience of experience in biopharmaceutical • GLP Clinical & Preclinical Bioanalysis • Recombinant Proteins & Glycoproteins development support across a wide range (PK, ADA, Nab) • Monoclonal Antibodies of product types. We provide regulatory- • Immunogenicity Studies led, phase-appropriate, tailored analytical • Antibody-Drug Conjugates program design with GLP / GCP / GMP • Analytical Programme Design • Biosimilars / Biobetters compliant laboratory services which • Structural Characterisation (ICH Q6B) • Peptides help you to navigate the challenges of • Physicochemical Properties (ICH Q6B) • Growth Factors development, regulatory submission, and • Biophysical Characterisation manufacturing. • Interferons • Comparability Studies With facilities located in India, Europe (UK, • Interleukins France, Switzerland) and the USA, our • Biosimilar Programmes • Vaccines strong scientific and technical leadership • Process Residuals Determination • Viral Vectors coupled with project management and • Product Related Impurities Determination • Oligonucleotide Therapeutics regulatory support, deliver a responsive, • Purity and Impurity Assessment flexible (bio)analytical resource, to drive your development and manufacturing • GMP Potency / Cell Based Assays programs forward. • Method Development & Validation • Extractables / Leachables • GMP Quality Control Testing • GMP Batch & Final Product Release Testing • ICH Stability Studies • Forced Degradation Studies • Drug Delivery / OINDP Expertise INTERTEK BIOPHARMACEUTICAL SERVICES

To achieve a well- Protein Characterisation characterised biologic, Protein therapeutics can present significant analytical challenges. Our characterisation we apply a strategic packages are tailored to your biomolecule analytical program and reflect the requirements of ICH Guidelines to meet the analytical challenges that is tailored to your of a ‘well characterized’ or ‘specified’ molecule. biological product. Programs encompass many different analytical techniques and provide information ranging from evaluation of physicochemical properties and structural features including primary, secondary and higher order structure and assessment of post-translational modifications. Programs also include determination of biological potency and assessment of purity /impurity profiles. We continually invest in advanced analytical instrumentation which allows us to deliver data with the highest sensitivity, accuracy and resolution.

Structural Characterisation Amino Acid Sequencing/ Sequencing studies and peptide mapping using a broad range of enzymatic or chemical digestion followed by Peptide Mapping: Mass Spectrometry (LC-MS/MS or MALDI-TOF MS). Amino acid composition: Pharmacopeia methods. Terminal amino acid Confirmation of N- and C-terminal sequences and evaluation of modifications and / or heterogeneity. sequence: Disulphide bridge Assessment of the degree and positions of both expected and mis-matched disulphide bridges by extended mapping: LC-MS/MS study and colorimetric test for free sulfhydryl groups. Glycosylation studies typically including levels of monosaccharides and sialic acid, N/O linked glycan profiling Carbohydrate structure: (NPLC, HILIC, IEX or CE-LIF), enzymatic digest and MALDI –TOF or LC-MS/MS. Physico-chemical Properties Molecular weight of intact proteins by MALDI-MS, ESI-MS and LC-MS supported by orthogonal techniques Molecular weight: such as MALLS and SDS-PAGE. Isoform pattern: Isoform and impurity studies using PAGE, SDS-PAGE, IEF, CE, HPLC. Extinction coefficient: Determination and Validated Extinction Coefficient studies. LC patterns: For ID, homogeneity, purity – HPLC, UPLC, SEC, RP HPLC, IEX, AEX. Spectroscopic patterns: CD, FTIR, 1D & 2D NMR, Fluorescence, UV-Visible. Electrophoretic patterns: CE(CZE), cIEF, CGE, SDS and NATIVE PAGE, Western Blot. Concentration: Lowry, BCA, Total AA, Total Nitrogen, Bradford. Aggregation studies: SEC (MALLS), DLS, Western Blot, CE, Gel Electrophoresis, SEM/TEM. Process Impurities: Residual host cell DNA by qPCR, Residuals (such as antibiotics, antifoaming agents). Potency Assays To support characterisation, stability, comparability testing and product release, for example, Complement- Cell-based Assays: dependent cytotoxicity (CDC), Antibody–dependent cell cytotoxicity (ADCC) and Neutralisation and Proliferation Assays.

3 Bioanalytical Expertise We apply our bioanalytical expertise and industry Large molecule bioanalysis Biomarker assays We have extensive experience in the We deliver discovery and clinical biomarker insight to design strategic development, validation, and sample analysis of solutions to support your precision medicine and efficient bioanalytical quantitative and qualitative Good Laboratory strategy enabling you to better predict Practice (GLP) and Good Clinical Practice (GCP) the long-term safety and efficacy of your programs immunochemistry and LC-MSMS expertise in products. Our dedicated biomarker team has support of pre-clinical and clinical development expertise in the qualification and validation programs in full regulatory compliance (FDA, of biomarkers using ELISA, and ECL platforms EMA and OECD GLP regulatory standards). (including multiplexing, prototypes and custom multiplexing and Luminex) in multiple matrices and anticoagulants. Our bioanalytical experts are highly experienced in developing and validating assays for • Quantitative Ligand Binding Assay pharmacokinetic (PK), toxicokinetic (TK), Capabilities pharmacodynamic (PD), immunogenicity, • Quantitative ELISAs for Proprietary efficacy and safety biomarker studies with Compounds diverse immunochemistry technologies, • Immunogenicity Studies radioimmunochemistry (RIA) and functional • Neutralization Cell-based Assay assay platforms. Development, Validation and Sample We offer comprehensive bioanalytical services Analysis to support complex products including proteins, • Radioimmunoassays (RIA), Enzymatic synthetic peptides, humanized monoclonal Assays antibodies, antibody drug conjugates, conjugated drugs, growth factors, hormones • Fluorometric Assays, Luminescence Assays and cytokines. Additionally, we offer industry • Biotinylations & Ruthenium Labeling unique capabilities, for example, quantitative, • Mode of Action Studies high resolution NMR which is an ideal tool for • Bioanalytical LC-MS/MS for Biologics bioanalysis of PEGylated moieties. • Affinity Interactions by SPR • NMR approaches for PEGylated Biomolecules • Biomarker Assays

CASE STUDY Specialist Expertise Novel Approaches for An Enzyme Activity Assay Biosimilar services Monoclonal antibodies A client desired an activity assay for a Our biosimilar thought leaders apply a strategic Our experience spans recombinant monoclonal PEGylated enzyme for which a commercial approach to developing biosimilarity data antibodies and related products such as colorimetric assay was available. The with programs that provide highly relevant biosimilars, fusion proteins, Fab-fragments and colorimetric assay did not meet the early stage characterisation and late stage Fc fragments and antibody drug conjugates performance criteria for regulated work. An comparative data. These programs begin (ADCs) throughout the product lifecycle with immunoassay was then developed but was with extensive structural and functional a focus on monitoring relevant critical quality subject to significant matrix effects. characterization of both the proposed biosimilar attributes (CQAs), assessment of the impact of Our Solution product and the reference product and process changes on physicochemical properties Development of a replacement assay was evaluate and compare all pertinent features and structure, the presence of product-related complicated by endogenous substrate and of the biosimilar product and are based on the impurities or process-related impurities. enzymatic product.To overcome this, the criteria outlined in ICH Q6B. This analytical • Characterisation (Q6B) specificity of LC-MS/MS was employed and characterization serves as the foundation of • Primary Sequence Confirmation an activity assay developed using a stable a biosimilar development program. Programs (Peptide Map) labeled substrate which produced a labeled encompass many different analytical techniques product, which could be differentiated from and provide information ranging from evaluation • N/C terminal Sequence, Disulphide Bridge the endogenous analyte. of physiochemical properties and structural Mapping and PTMs Benefit Delivered to our Client features including primary, secondary and • Carbohydrate Structure A enzymatic activity LC-MS/MS assay was higher order structure and assessment of • Aggregation Studies post-translational modifications and cell-based developed and successfully validated to • Higher Order Structural Studies regulatory standards. The method was used assays to determination of biological potency • Aggregation Studies in multiple pharmacokinetic studies and with the goal to show that the molecule is enabled the client to move forward with structurally and functionally highly similar to • Heterogeneity, Purity and Variants their drug development program. a reference product and so is anticipated to • Potency behave like the reference product. INTERTEK BIOPHARMACEUTICAL SERVICES Protein Analytics

Post Translational Modifications (PTM) Potency assays Process residuals determination We apply our knowledge of anticipated PTM Potency bioassays are an integral part of release Our process residual analysis services apply and potential PTMs based on your product’s and stability testing. Intertek’s experienced a range of highly specific and sensitive specific structural characteristics to design specialists perform cell based potency and techniques to residual cell substrates (e.g., host strategic analytical programs that meet the ligand binding assays which can be performed cell proteins, host cell DNA), cell culture (e.g., requirements of regulatory expectations and to GMP for regulatory submissions or as a non- inducers, antibiotics, or media components), or industry guidance such as ICH Q6B. Using regulatory study under high quality standards. chromatographic media used in purification, enzyme or chemical digestion coupled with Glycosylation solvents and buffer components to offer robust highly sensitive liquid chromatography tandem Glycosylation studies are designed to be quantification supporting process validation, mass spectrometry (LC-MS/MS) and a range of product specific, however, these typically monitor the batch to batch variation and support chromatographic techniques we can detect and include determination of the levels of neutral GMP lot release. For trace antibiotics such as identify a range of PTMs. and amino monosaccharides as well as sialic Kanamycin, Tobramycin, Gentamycin, Amoxicillin, • Deamidation • Methylation acids, assessment of glycoform distribution Chloramphenicol, we apply highly selective approaches such as tandem mass spectrometry • Glycosylation • Oxidation and glycan structure elucidation. Multiple technologies are applied to the determination (LC-MS/MS) with Multiple Reaction Monitoring • Phosphorylation • Mis-matched S-S (MRM). We also apply MS techniques to the bridges including selective enzymatic cleavage and • Acetylation MALDI-TOF Mass spectrometry HPLC, HILIC, determination of surfactants and antifoams • Alkylation • Truncation IEX or CE-LIF, to provide the level of structural agents which typically include Triton-X, Tween 20, Tween 80, Pluronics and polyglycol P2000. • Sulfation • N/C-terminal information required. Modifications • Glycylation Product related impurities • Kanamycin • Tween determination • Tobramycin Biophysical characterisation • Pluronics Our scientists perform detailed characterization Our biophysical characterisation team applies a • Gentamycin • Polyglycol P2000 using a diverse range of technologies which wide range of techniques such as to interrogate • Amoxicillin include chromatographic, electrophoretic, • Trace metals the biophysical behavior of your molecule. We mass spectrometry (MALDI-MS, LC-MSMS) • Chloramphenicol examine product’s higher-order structure (HOS), • Downstream- and spectroscopic (IR, NMR and fluorescence including secondary and tertiary structure using • Methotrexate derived impurities approaches) to determine truncated forms Circular Dichroism (CD), FTIR, Fluorescence, 1D • Glutathione and other modified forms such as deamidated, • Leachables / 2D High field 600MHz NMR studies (NOESY isomerised, mismatched S-S linked, oxidised or • Dithiothreitol from process & TOCSY), aggregation (using SEC–MALS, DLS, altered conjugated forms (e.g., glycosylation, equipment SV-AUC), oligomerization and degradation of • IPTG phosphorylation). For truncated forms, we use the drug substance. To support formulation • Triton-X • Particulates an enzyme or chemical cleavage of peptide development we study proteins in the buffer followed by characterisation by chromatography and in the presence of additives / excipients of (e.g. HPLC) or SDS-PAGE approaches. We interest. To support IND or NDA submissions also perform peptide mapping to yield useful we assess spectroscopic, thermodynamic and information about the variant. hydrodynamic properties. Development Support

ICH stability testing testing. Release tests can be developed and With dedicated ICH storage stability facilities validated in-house, transferred from clients or that are integrated with advanced analytical include Pharmacopeia Monograph testing. capabilities, we can offer stability-indicating Formulation development method development and validation alongside Formulation of biopharmaceuticals for inhaled efficient stability programs that focus on the or nasal delivery allows for a more convenient critical quality attributes OF your molecule. method of administering compounds Comparability studies systemically, and also allows direct targeting of The FDA and EMEA require demonstration the respiratory system, however, this brings new of comparability compliant with ICH Q6B and challenges. Our experts integrate formulation Q5E guidelines. Intertek has undertaken many with tailored analytics that focuses on the detailed comparability studies since 1999. There impact of formulation on the safety and efficacy is a regulatory expectation that “orthogonal” of the drug whilst monitoring aggregation, approaches are applied such that conclusions fragmentation, deamidation, hydrolysis, for key quality attributes are based on multiple oxidation, isomerisation, succinimidation, approaches. deglycosylation, disulphide bond formation/ QC release testing breakage and other crosslinking reactions. Important for both bulk lots and finished products, our suite of release tests includes assays for concentration, purity and quality

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