Protein Therapeutics

RSSL’s multidisciplinary team of experts use cutting edge instrumentation and dedicated facilities to provide comprehensive support for your protein therapeutics. 

 

Our tailored analytical solutions are designed to help clients in fully understanding their protein based therapeutics at all stages of of their journey - from development to batch release testing

 

Therapeutic protein drugs are complex and heterogeneous molecules often produced from bacterial, mammalian or insect cell lines. This key class of medicines is used to treat a wide range of conditions, such as cancers, autoimmune diseases, inflammatory disorders, infections, and genetic conditions, highlighting the need for rigorous and reliable testing.

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Our services

 

At RSSL, we have extensive expertise in a broad range of peptides and protein therapeutic measurements using conventional and advanced analytical methods including bioactivity measurements.

 
Our multidisciplinary team of experts will guide you through the analytical steps needed to fully characterise your biomolecule, understand your raw material and ultimately accelerate your drug product development.


With services designed to help you meet the characterisation, comparability and release testing requirements as outlined in the International Committee Harmonisation (ICH) guidelines such as the ICH Q6B and the ICH Q5E, our history in current Good Manufacturing Practice (cGMP) makes RSSL the analytical partner of choice for biologicals.

 

Our expertise incorporates monoclonal antibodies, bispecific antibodies, enzymes, cytokines, antibody-drug conjugates and biosimilars through services such as:

 

  • Physical chemical characterisation
  • Impurity analysis
  • Activity and potency 
  • Stability   
  • Pharmacopeia testing
  • Environmental and sterility testing   
  • Raw material testing

 

Physical Chemical Characterisation

 

Our comprehensive physical and chemical characterisation services for protein therapeutics are fully aligned with ICHQ6B, EMEA, and FDA guidelines. With state-of-the-art analytical techniques, we ensure accuracy, compliance and deep insights into the molecular properties of clients protein-based products. 

  • Molecular weight determination is carried out using high-accuracy mass spectrometry analysis. Proteins are analysed in their intact form by reverse phase chromatography or size exclusion chromatography coupled with mass spectrometry used to determine the molecular mass of the protein for greater accuracy and precision.

     

    Complementary electrophoretic techniques such as SDS-PAGE and CE-SDS with UV or fluorescence detection provide detailed verification of protein profiles. Additionally size exclusion chromatography (SEC) equipped with UV or fluorescence detection supports molecular mass assessment distribution to determine the presence of aggregates. 

  • We conduct chromatographic and electrophoretic profiling using techniques such as size exclusion chromatography (SEC), ion exchange chromatography (IEX), reverse-phase chromatography (RP) and capillary electrophoresis (CE-SDS, CE-IEF, and CZE).

     

    These methods provide clients with detailed insights into heterogeneity, isoform distribution and purity of the protein and can be used at various stages of the product life cycle alongside conventional methods such as SDS-PAGE and Western Blotting.

  • We determine protein quantification using European Pharmacopoeia (EP) and United States Pharmacopeia (USP) methods, ensuring compliance with global standards. Measurements are carried out using colorimetric assays to quantify the total amount of proteins once the extinction coefficient and the amino acid quantification is established.

  • Protein sequence confirmation and identification of post-translational modification (PTM) analysis is performed at RSSL by proteolytic digestion followed by liquid chromatography-tandem mass spectrometry (QToF or Orbitrap).

     

    This approach enables accurate confirmation of the protein sequence including N- and C-terminal sequence confirmation, location of disulfide bonds and identification/location of post-translational modification (including oxidation, deamidation, acetylation, ubiquitination and glycosylation). A comprehensive understanding of protein modifications provides critical information to determine the functionality and stability of the product.

  • RSSL carry out the extinction coefficient of proteins through UV/visible absorbance measurements using solutions with known protein content accurately quantified by amino acid analysis.

  • Higher order structural analysis is performed using far- and near-UV circular dichroism (CD) and Fourier-transform infrared spectroscopy (FTIR) which provides insights into the structural integrity. RSSL also performs aggregation studies using techniques such as size exclusion chromatography and dynamic light scattering (DLS).

     

    We provide additional methodologies including multi-angle light scattering (MALS), sedimentation velocity analytical ultracentrifugation and differential scanning calorimetry (DSC) to provide clients with vital additional assessments of aggregation ensuring a thorough understanding of protein under various conditions.

 

Impurity Analysis

 

Impurity analysis is a critical aspect of protein therapeutic quality control. At RSSL, we offer a broad range of services addressing residual solvent analysis, process related impurities and product related impurities of biopharmaceuticals in line with the ICH Guidelines such as ICHQ1A, ICHQ3A, ICHQ3B, ICHQ3D and ICHQ6. 

  • Process related impurities are residues from the manufacturing process which include cell substrates such as host cell proteins and host cell DNA. Impurities can also include cell culture derived impurities such as antibiotics and cell culture media residues or downstream derived impurities such as enzymatic/ proteolytic agents, reducing agents, inorganic salts, solvents, nitrosamines and leachables

     

    RSSL reputation for cGMP regulatory compliance, implementation of pharmacopeia monographs and comprehensive advanced analytical and bioanalytical methods provide the ideal setting for investigative and quantitative studies of process related impurities in bio-products and for generating data for regulatory approval.

     

    RSSL process related impurity techniques include liquid chromatography and gas chromatography, mass spectrometry (MS), nuclear magnetic resonance (NMR), inductively couple mass spectrometry (ICP-MS), polymerase chain reaction (PCR), enzyme linked immunosorbent assays (ELISA) and enzymatic assays. Our expertise in chemistry, elemental analysis, biochemistry, molecular and cell biology help define the best strategy to meet regulatory requirements and accelerate the development of client’s products. 

  • Molecular variants, such as product related impurities arise during the manufacture and/or storage. These do not have the same properties as the desired product. Examples of product-related impurities can include truncated and other modified forms, aggregates, precursors and degradation products

     

    We work with clients to provide tailored analytical workflows that meet client’s needs, using chromatographic, electrophoretic and mass spectrometry methods. 

     

    Deamidation, oxidation, mismatched disulfide bridges and altered post-translational modifications such as glycosylation or phosphorylation are separated, quantified and identified using chromatographic, electrophoretic and high-resolution mass spectrometry methods. 

     

    Due to extensive capability, RSSL are able to offer the bespoke service of native size exclusion chromatography coupled with high resolution mass spectrometry additionally to other methods such as UV, FLD and MALS detection. This service monitors complex aggregate integrity and identity.  

Activity and Potency

 

Potency assays are essential for evaluating the biological activity of drug substances and products throughout the development lifecycle. Ligand binding assays which use the specificity of an antibody to its antigen are commonly used to quantify the amount of drug substance or product in a sample. These assays often serve as surrogate potency or activity methods.

 

As a therapeutic progresses towards the last phases before commercialisation, a cell-based potency assay becomes a regulatory requirement to measure the drug's quantifiable biological response and active content. Utilising cutting edge facilities, RSSL can develop both cell-based potency assay and ligand binding assays to determine activity and potency of bio-product at early stages of drug development. This data supports stability and formulation testing and forms part of the GMP batch release testing requirements. 

 

RSSL also specialises in developing bespoke cytotoxicity assays, including USP-87 assays and proliferation assays. These assays are tailored for anticancer therapeutics, mitogenic drugs ensuring reliable potency evaluation. To further reinforcing RSSL as the partner of choice for activity and potency testing, we have strong collaborative partnerships with antibody and cell line producers. 

 

 

Stability Testing

 

With extensive expertise and state of the art storage facilities, RSSL routinely carries both long-term and accelerated stability (6 months) tests under a broad range of conditions tailored to customer requirements. For long-term storage (12 months), samples are most frequently stored at -20 °C ± 2 °C or 5 °C ± 3 °C, with analysis at 3, 6, 9, and 12-month intervals. For the accelerated stability study (6 months), conditions of 5 °C ± 3 °C or 25 °C ± 2 °C / 60% RH are generally considered at the 3 and 6-month timepoints.

 

The testing analytical protocols for each stability study is agreed with each client. This includes the broad range of available analytical and biomolecular methods for characterisation and batch release testing. 

 

 

Pharmacopoeia

 

Our highly skilled team of analysts have decades of experience providing reliable, accurate and timely results in accordance with various major pharmacopoeia. Our testing capabilities encompass a wide range of techniques structured to support quality control throughout the pharmaceutical development and manufacturing process, from raw materials to finished products.

 

 

Enviornmental and Stability Testing

 

RSSL provide consultation and analytical services for environmental monitoring to enable manufacturers to accurately determine microbial bioburden. Also carried out by RSSL are microbial identification services which assist clients in defining root cause analysis and corrective and/or preventive action implementation.

 

This analysis enables clients to meet regulatory requirements such as Ph. Eur, JP and USP regulations reliably and cost-effectively for terminally sterilised or aseptic products. 

 

Get in touch to find out how we can transform complex chemistry into reliable results for you

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