Analytical Development

In addition, AS helps, along with our biologics Discovery R&D teams, to identify the final lead drug candidates by testing multiple candidates for quality, stability and developability/manufacturability, thus reducing the risks for the subsequent CMC development process. 

A typical developability study includes the following aspects: 

• Prediction of high-risk PTMs
• Titer level
• Aggregation tendency
• Thermal stability
• High-concentration solubility and viscosity
• Short-term stability (thermal, freeze-thaw, forced degradation)

• Our highly experienced analytical team supports our CMC development services across all process development functional groups, and also supports the investigations of GMP manufacturing issues.
• During development, a collection of compendial, physicochemical, biochemical and potency assays are carefully selected based on scientific consideration and the deep understanding of our platform processes and assays to monitor product quality attributes and support process development.

• Our highly skilled experts from the Mass Spectrometry, Forensic Analysis and Bioassay Centers of Excellence (CoE)  provide high-quality product characterization services for our customers.
• We are capable of comprehensive product and reference standard characterization on the molecule’s biochemical, biophysical and biological characteristics.
• For novel drugs, we can conduct head-to-head comparison on release and characterization test items between GMP and tox lots. For biosimilar drugs, we can perform extensive similarity assessment.
• For late-stage projects, we offer detailed characterization services to gain more insights into the product, such as charge variants and isoforms observed in forced degradation studies.

• Our AS team performs testing on non-clinical lot release and stability samples, while the GMP lot release and stability are performed by the quality control team.
• We have a comprehensive mAb-based platform testing panel for lot release. A customized  release panel is also available based on specific requirements from different filing regions or product characteristics and for non-mAb products.
• For each product, a specification will be established for GMP lot release, following phase-appropriate strategies. During early phase method development, specifications with relatively broad ranges will be established based on process development data, industrial experience and regulatory requirements. As the project enters a later phase, the specification can be narrowed based on statistical data from multiple GMP lots.
• Reference standard (RS) release also follows a life-cycle management strategy, including protocol-driven RS generation, qualification and annual re-qualification, routine management, and the bridging of interim, primary and working reference standards.

• We have a dedicated stability team to handle non-GMP and GMP  stability programs, adhering to the same set of management practices and high operational standards per SOPs and protocols. 
• The stability program operates based on the agreed upon stability protocol. Its testing process is controlled by our LIMS, which covers stability program configuration, sample pulling schedule management, and sample and testing tracking.
• We provide basic stability programs for long-term, accelerated and stressed conditions, with well-designed time points to span across a long timeframe for establishing product shelf-life. A customized program is also available to suit customer’s clinical study timeline and product quality monitoring purposes.

• Our analytical team is dedicated to preparing a well-structured Quality section of the Common Technical Document (CTD) for applications that will be submitted to regulatory authorities. The data integrity verification process will be completed before the dossier is finalized.

• Critical Quality Attribute (CQA) analysis and Forced Degradation Study (FDS) are conducted to ensure the design of regulatory compliant stability programs​
• Assay qualification for in-process samples to support PC (Process Characterization) and PPQ (Process Performance Qualification) campaign
• Transfer to Quality Control group for Assay Validation to support late phase and commercial products
• Comparability studies ensure robust release assays used for clinical stage and PPQ product batches.
• Process-specific HCP (Host Cell Protein) services along with fractionation and characterization analysis are available for product variants

We provide a comprehensive range of assays to detect process-related impurities. Listed below are examples of impurity detection assays previously developed and validated by the WuXi Biologics Process Related Impurity Detection Center of Excellence. This list is not all-inclusive of all assays developed by the Center of Excellence. Contact us to learn more.

We follow appropriate testing procedures that comply with CP, USP and/or EP for each integrated project depending on its filing region.

• General assay:

• • Appearance
  • Color
  • Clarity
  • Visible Particles
  • Subvisible Particulate Matter
  • pH
  • Osmolality
  • Extractable Volume
  • Water Determination

   

• Biosafety: 

• • Sterility
  • Bioburden
  • Endotoxin

For information on our compendial mycoplasma assays and other non-compendial biosafety tests (e.g., adventitious virus), click here.

We offer biochemical analysis services for the identification, detection and quantitation of process-related impurities/contaminants, as part of an integrated package and as standalone projects with rapid turnaround. We offer platform methods and our team is experienced in managing biochemical methods throughout the lifecycle of development/optimization, transfer, qualification and sample testing. The following list illustrates some of our platform biochemical analysis capabilities.

• Impurity：

• • Residual Host Cell Protein  (HCP), Residual Protein A, Residual DNA by qPCR
  • Quantitative ELISAs for impurities including individual HCP, hormones and enzymes
  • DNA product-related methods (quantity, identity and impurities) 
  • Gel Electrophoresis
  • Western Blot & 2-D Western Blot
  • Gene Expression by qPCR

• Mass-Spectrometry (MS)-based Analytical Testing：

• • Mass-Spectrometry-based approaches (MS) are implemented throughout the entire mAb development and production process, providing information on primary structure and Post Translational Modifications (PTMs) with tests such as Mass Analysis, Peptide Mapping, and N-glycan Identification.
  • Mass Analysis: intact/reduced mass, glycosylated/deglycosylated reduced mass
  • Peptide Mapping: sequence coverage, PTMs, C-/N- terminal sequencing, disulfide bond analysis, peptide mapping for identification 
  • Glycan Analysis: N-/O-glycosylation profiling and site confirmation, glycopeptide analysis
  • Advanced techniques: sequence variant analysis, impurity characterization, mutation and rare PTMs ID, individual HCP ID and quantitation
     

• HPLC/CE based:

• • The purity of drug substance and drug product is assessed by a combination of analytical procedures. Our analytical team is dedicated to providing platform procedures and developing customized procedures for unconventional molecules like fusion proteins and bispecific antibodies.
  • Aggregation: SEC
  • Fragment: reduce/non-reduced CE-SDS
  • Charge Variants: cIEF, AEX, CEX
  • ADC related: HIC

We provide biophysical techniques for probing the higher-order structural integrity and for full product characterization. The tests include secondary and tertiary structure analysis, thermal stability, and molecular weight distribution.

• Biophysical：

• • Far UV CD / FTIR
  • Near UV CD
  • DSC
  • SEC-MALS / AUC
  • Fluorescence
  • Free sulfhydryl analysis

Assessment of the biological properties constitutes an essential step in describing the specific ability or capacity of a product to achieve a defined biological effect. We offer biochemical assays, cell-based assays and other ligand/receptor-binding assays to measure the biological activity and explain the correlation between the expected clinical response and the biological assays.

• Biological:

• • • Fab functional characterization
    • Antigen-binding assays (ELISA, SPR, FRET, Alphascreen, etc)
    • Homogenous cell-based binding assay (ECL, FACS, etc)
    • Anti-proliferation assays
    • Cellular signal transduction/apoptosis/killing assays
    • Neutralization assays
    • Adhesion assays and migration assays
    • Antigen-specific immune response and its regulation: T-dependent human and murine responses

   

• Fc functional characterization

• • FcγR and FcεR binding assays (ELISA, SPR, FRET, Alphascreen, etc)
  • FcRn binding assays (ELISA, SPR)
  • C1q binding assays
  • Complement dependent cytotoxicity (CDC)
  • Antibody-dependent cell-mediated cytotoxicity (ADCC)

   

• Enzymatic Activity Assays

Following the unique properties of the molecule, WuXi Biologics Analytical Sciences (AS) team offers a full set of analytical methods leveraging physical-chemical, biological, higher-order structural, and protein structural analytical technologies. AS has extensive experiences in analytical method development for different types of proteins, including but not limited to monoclonal antibodies, fusion proteins, antibody drug conjugates (ADCs), recombinant proteins and bispecific antibodies.

For the DS/DP release assays, we provide analytical method qualification/validation following ICH Guideline Q2(R1), FDA Guidance for Industry, PDA Technical Report No. 57, and USP General Chapter <1225>&<1226>. Assays to be qualified/validated include product-specific methods, process-related methods and microbiological methods.

We offer method transfer services to the receiving lab for analytical methods including but not limited to in-process testing assays, DS & DP release assays and characterization assays.

We offer method transfer services to the receiving lab for analytical methods including but not limited to in-process testing assays, DS  and DP release assays and characterization assays.