SGS Biopharmaceutical Services

Pharmacokinetic (PK) and Toxicokinetic (TK) studies are usually conducted at the discovery or drug candidate selection stages of the drug development lifecycle. PK and TK studies provide insight into fate of a drug upon administration.

In order to support PK and TK studies of biopharmaceuticals, a wide range of immunochemistry platforms such as ELISA, EIA, ECL, Luminex, LC-MS and methods combined with scientific expertise are available. Services include:
o Pre-clinical and clinical support
o Custom Immunoassay Method Development
o Method Transfer
o Method Optimization
o Method Validation - Validation criteria follow guidelines from the FDA and the Crystal City Conference. All validations are tailored to specific program requirements.
o Routine Testing
o SGS offers validated bioanalysis methods that are ready for use

Dried Blood Spot Analysis
Dry Blood Spot (DBS) technology has several advantages for toxicokinetic (TK) and pharmacokinetic (PK) analysis, including: reduced blood sampling collection, absence of post-collection processing, low biohazard risk, fewer required facilities for sample shipment and storage. Furthermore, sample analysis can also be simplified.

Host Cell Protein (HCP) are process-related protein contaminants / residuals present in low levels during production of biological drug products derived from biomanufacturing processes. These proteins have to be removed during purification as presence of these impurities may potentially compromise the safety and efficacy of a biotherapeutic product, which could ultimately have a negative impact on patients. Thus, HCP analysis comprises of an important component in the safety evaluation of biopharmaceuticals.

At SGS we offer Host Cell Protein (HCP) Analysis services. Methods can be transferred, validation and used for routine testing e.g. during GMP release tests.

Biomarkers are indicators of a biological state or condition – normal or diseased. Biomarker analysis is important during the developing of a drug to help with:
a. Understanding physiological impact of a drug
b. Understanding the pharmacological mechanisms of a drug
c. Uncovering potential new drug targets
d. Ultimately understanding the safety and efficacy of a drug

Thus, biomarker assays have increasingly become an integral part of clinical development as biomarkers can be used as early indication of drug efficacy.

SGS offers biomarker analysis of a variety of samples from pre-clinical to human clinical trial samples. Our biomarker testing services support the pharmaceutical and biopharmaceutical industry by developing, validating and routinely testing relevant biomarkers in biological fluids / matrices using a variety of platforms including:

• ELISA – sandwich ELISA, competitive assays, singlex
• Multiplexing - Luminex (bead-bases suspension) and Electrochmiluminescence (ECL) based technologies to analyze multiple analytes simultaneously

Biomarker assay capabilities include method development, optimization, transfer, fit-for-purpose validation and routine testing. Our team of scientists in our European and USA Good Laboratory Practice (GLP) facilities can support your global clinical development programs.

In collaboration with the clinical team, SGS has actively pursued innovative biomarker assay development and validation. Our biomarker bioanalytical services cover a range of therapeutic areas to include:
• Cardiovascular
• Central nervous system (CNS)
• Metabolic disorders
• Inflammation
• Thyroid disease
• Oncology
• Sexual health
• Bone disease
• Allergy and respiratory

SGS offers a wide range of immunoassay and cell-based assay services. The following services are offered:

• Potency Assays
• Immunogenicity studies
• Residual analysis such as Host Cell Proteins (HCP), Host Cell DNA (HCD), Protein A, etc.
• Anti-Drug Antibody (ADA) Assays
• Neutralizing Antibody (NAb) Assays
• PK Assays
• Biomarker Assays
• ELISA, EIA, ECL Assays
• Quantitative and Qualitative Assays

In collaboration with our clients, SGS can develop, optimize, transfer, validate custom assays, conduct routine testing to meet the specific needs of our clients. Typical platforms that are often used are:

• Multimode plate readers - absorbance, fluorescence intensity / polarization, time-resolved fluorescence, luminescence.
• Meso Scale Discovery (MDS)
• Luminex
• FACS

Assessment of Neutralizing Antibodies (Nabs) during an immunogenicity study is important because they bind to the epitope(s) within the active site of the therapeutic molecule inhibiting its biological activity. The most serious adverse effect of Nabs is to not only neutralize the therapeutic protein but also cross-react with and neutralize the endogenous protein.

The following includes SGS’s services and expertise on NAb assays:
o Neutralizing Antibody Assay
 Assay Design: selection of a cell line, assay endpoint, and positive control antibody
 Assay Formats: Direct Nab assay, Indirect Nab assay, Matrix Interference assay (confirmatory assay)
 Assay Optimization: Selection of a drug concentration, sample matrix dilution assay robustness, assay incubation conditions.
 Assay Qualification: Characteristics of the drug product standard curve, positive control antibody curve, assay cut points for the Nab assay, assay cut point for the matrix interference assay, assay sensitivity, and assay characterization.
Neutralizing antibody assay uses a tiered approach where study samples at appropriate time points are initially analyzed using an immunoassay for the presence of anti-drug product antibodies (binding antibodies). Samples yielding a positive result in the immunoassay are subsequently tested for anti-drug neutralizing activity using an in-vitro cell-based assay or a non-cell based competitive ligand binding assay.
Confirmatory assays are used to determine a true neutralizing antibody response from other interfering factors that may be present in the serum sample that could mimic a Nab effect in the absence of Nabs. This is used to detect drug specific neutralizing antibodies.
Cell based assay (direct or indirect) is preferred due to its physiological relevance, but non-cell based assay (direct or indirect) can also be utilized if cell based assay is not available.
The different types of technologies that can be used are FACS, Imaging, Branched DNA technology, ELISA, and ECL.
Regulatory agencies recommend the usage of Immunogenicity testing of drug products during early and late stage development to commercialization.

SEC-UV

High order structure evaluation
Aggregation: SEC-MALS, SV-AUC, DLS, SEC-UV, Gel electrophoresis
Secondary/tertiary structure: FTIR, CD, Fluorescence

Generating a stable environment for a biopharmaceutical drug product is a critical step for ensuring a long product shelf life. In order to maintain potency and activity, some of the most critical parameters to maintain are protein conformation (tertiary or 3-dimensional [3-D] structure), folding (secondary structure) and proper subunit association (quaternary structure). Collectively, these are known as higher-order structure (HOS) and can be highly influenced by the formulation environment, manufacturing process and storage conditions. However, analytical monitoring of the protein conformation has been frequently omitted because of time constraints, the complexity of techniques commonly employed and the inherent lack of sensitivity to subtle changes shown by some biophysical methods. The use of traditional biophysical techniques, combined with more sensitive orthogonal approaches, provides a comprehensive view of higher order structure in biopharmaceutical formulation.
Biophysical analysis has become increasingly important in the characterization of biopharmaceutical drug candidates and the analyses used may be described as covering a mixture of disciplines, essentially being the use of physical techniques to investigate and characterize biological systems. A wide range of analyses are performed within this area for the elucidation of 3-D protein structure, including the application of spectroscopic, thermodynamic and hydrodynamic techniques, such Circular Dichroism (CD), Fourier transform infra red (FT-IR) spectroscopy, Differential Scanning Calorimetry (DSC), Intrinsic and Extrinsic Fluorescence, Dynamic Light Scattering (DLS), Size Exclusion Chromatography Multi-Angle Light Scattering (SEC-MALS) and Sedimentation Velocity Analytical Ultracentrifugation (SV-AUC). Analysis of these properties can provide valuable information on secondary, tertiary and quaternary structures of protein molecules such as monoclonal antibodies, and can provide evidence that the molecule maintains conformational stability.

Plasmid Copy Determination

SGS laboratory experts have extensive knowledge and experience in pharmaceutical method development and validation for raw materials, APIs, finished products, cleaning and bioanalytical testing.
We offer development and documentation of analytical protocols and reports for proprietary and non-proprietary test methods and manufacturing processes. This is conducted in compliance with the Notes for Guidance Validation of Analytical Procedures, Definitions and Terminology and Validation of Analytical Procedures, Methodology by the ICH (Q2A, Q2B) and FDA guidelines.
Once a method is validated, it may require transfer. Method transfer may involve comparative testing, co-validation between two sites (lab-to-lab), complete or partial revalidation and comprehensive documentation (transfer plan, protocol, report).
Whether SGS’s Life Science Services is the developing or the receiving laboratory, we can assist you with your method transfer requirements.

Method Feasibility (Pilot Studies)

Method feasibility will be performed as an initial evaluation of the fit for purpose status of the analytical methodology or as an initial evaluation of the fit for purpose status of SGS in-house analytical methodology applied to the client’s product. Included in method feasibility is ordering of materials and reagents, as well as minor non-method altering optimization to ensure compatibility with SGS Life Science Services technology and compliance systems. When acceptance criteria have been defined by the Sponsor, the Pilot Studies will incorporate some assessment of these acceptance criteria. On the completion of feasibility assessment, a GO/No-GO decision will be communicated prior to proceeding to method pre-validation, qualification or validation. Any significant development or optimization beyond the scope of the feasibility phase may be subject to additional fees. Feasibility or Pilot Studies will not be protocol driven. Depending on the nature of the test, reports may be issued.

Method Optimization

Based on the outcome of the feasibility phase of the project, methods requiring additional optimization may be optimized to assure that the fit-for-purpose status of the method is met. In the circumstance where optimization is required, the optimization strategy and pricing will be discussed and agreed upon by both organizations prior to initiation of the work.

Method Verification (Applicable only to compendial methods)

Verification protocols will be written by SGS Life Science Services and will include all method verification parameters and acceptance criteria. The protocols will be approved by SGS Life Science Services QA and the client before execution. A draft verification report will be presented to the client for review and comment prior to submission to SGS QA. Once the client’s comments are received they will be reviewed, discussed if necessary, and incorporated before issuance of a final report. SGS Life Science Services QA and the client will approve the final report.

Method pre-Validation/Qualification
Method pre-Validation/Qualification will be performed on non-compendial methods when acceptance criteria have to be defined. The pre-Validation/Qualification will be protocol driven and will be performed under cGMP. During pre-Validation/Qualification, a selection of Method Validation parameters outlined in ICH Q2(R1) will be assessed in order to define appropriate acceptance criteria to be used during the Method Validation. The set of parameters to be assessed may be discussed and agreed upon by both organizations prior to initiation of the work. Qualification Reports will be issued.

Method Validation
Method Validation will be performed as per requirements outlined in ICH Q2(R1). The protocols will be approved by SGS Life Science Services QA and the client before execution. On completion, reports will be presented to the client for review and comment. Once the client’s comments are received they will be reviewed, discussed if necessary, and incorporated before issuing a final report.

Method Transfer (Applicable only to validated methods)

Provision of the Validation Report is highly recommended for efficient and successful transfer. Transfer protocols will be written by SGS Life Science Services and will include all method Transfer the client before execution. A draft transfer report will be presented to the client for review and comment prior to submission to SGS QA. Once the client’s comments are received they will be reviewed, discussed if necessary, and incorporated before issuance of a final report. SGS Life Science Services QA and the client will approve the final report.

Our experts are highly experienced in both implementing client assays and developing them de novo. Our laboratories in France and Belgium are both GLP-certified and routine audits from the local agencies have proved that our laboratories consistently attain the highest standards in GLP compliance. In these facilities, we provide bioanalytical support for approximately 100 pre-clinical studies per year.

Our In Vitro Assay Service and Cell-based Bioassays Services include:
Models:

Monolayers: Customized sourcing of primary fibroblast or keratinocyte cultures
Human and animal cell lines (L929, Vero, IMR-90, MRC-5, CHO, HL-60, etc.)
3D skin model (epidermis – full thickness)
3D human corneal epithelium
3D human lung epithelium model
CACO-2 (human small intestine model)
Isolated human blood cells (PBMC, lymphocytes, monocytes, etc.)

Study types:

Barrier and metabolic functionalities as in vivo
Percutaneous absorption and metabolism, skin irritation, sensitization, corrosivity tests, phototoxicity, wound healing
Prediction of eye irritation potential of chemicals (alternative for in vivo Draize test)
Intestinal absorption and metabolism
Medical device cytotoxicity
Permeability of drug substances
Toxicity of API or excipients
API receptor binding assays
Wound healing
Physiological effect on cell cultures
Biomarkers synthesis and release
Bioassays for neutralizing antibodies
Cytokine production
Phagocytosis/Opsonization
Study of Intracellular Enzyme Activity (COX, MPO, etc.)
Study of Signal transduction (STAT-1, p38 MAPK,P44, AKT/PKB, etc.)
Immune cell activation and toxicity study types:
Biomarkers (cell receptor activation & signaling)
Phagoctosis of neutrophiles/macrophage
Opsonization
Phototoxicity
Flow cytometry: BD FACSCanto II (Immunophenotyping/Immunotox)
Immunogenicity of vaccines
Cell-mediated immunity

R&D Stability
Forced degradation studies
cGMP Stability

Product safety

• Immunogenicity testing
• Microbial testing

Product activity - enzymatic activity

Intact Molecular Weight (MW) or Molecular Size

For all compound classes, from small organic molecules to intact glycoproteins, the intact molecular weight is a key characteristic in its identity. Mass spectrometry offers accuracy of measurement over a wide molecular weight range, with minimal, often sub-picomole, sample consumption. Furthermore, any difference between theoretical and measured mass may be indicative of potential modifications.
Experience and our wide range of mass spectrometry instruments allows SGS Life Science Services to choose the most appropriate ionization technique for the compound, from MALDI-MS, ES-MS to an LC-MS experiment for complex mixtures. At higher resolution, measurement of molecular weight to within a few ppm, often to four or five decimal places by “accurate mass measurement”, can aid elucidation of the elemental composition of a compound.
Size exclusion chromatography (SEC-UV), gel and capillary electrophoresis and other methods such as SEC-MALS, DLS and AUC, are also available as alternative techniques to estimate molecular size.
Molecular weight analysis is a key part of the ICH Q6B guidelines for characterization and confirmation of biopharmaceuticals in support of new marketing applications.

METHODS

ESI-MS
LC-MS
MALDI-TOF MS
DLS
SEC-MALS
SEC-UV
SV-AUC
Gel electrophoresis

Isoform pattern: cIEF, iCE or IEX

Glycosylation site analysis

Monosaccharide composition analysis is used in the analysis of glycoproteins and carbohydrates to determine the identities and quantities of the various monosaccharides present. This information can be used at all stages of drug development as a method of determining the type of glycosylation (N-linked and/or O-linked) and the extent to which glycosylation has occurred.
Sialic acids are a class of monosaccharide of particular importance. Being located at the end of the antennae, they are considered as one of the major “capping” groups for N- and O-glycan structures.
Because of their location on the glycan moieties and therefore, their exposure to the external environment, sialic acids play a crucial role in the glycoprotein/glycopeptides function, clearance or binding.
A powerful technique in glycoprotein analysis, sialic acid analysis can also be used for:
• Screening of glycoproteins manufactured using different cell lines
• Demonstration of batch to batch consistency for release or during change in the manufacturing process
• Demonstration of similarity between a reference material and a biosimilar

Sialic acid analysis is most commonly performed using chromatographic techniques such High Performance Anion Exchange Chromatography with Pulsed Amperometric Detection (HPAEC-PAD).
Sialic acid analysis is a key part of the requirements laid out in the ICH Q6B guidelines for characterization and confirmation of biopharmaceuticals in support of new marketing applications.

METHODS
HPAEC-PAD, CE-LIF, HPLC-FLD

The ability to sequence the C-terminal region of a protein forms a critical part of a protein characterization package. C-terminal microheterogeneity frequently arises as a post translational modification, hence identification of the C-terminus and any truncated versions or 'ragged ends' is key in assessing the 'intactness' of a protein biopharmaceutical.
Unlike N-terminal sequencing, there is no fully reliable automated instrumental technique available. Confirmation of C-terminal sequence can usually be achieved using a combination of peptide mapping strategies and ES-MS/MS sequencing, combined with enzymic digestions and/or molecular weight analysis. Experience enables SGS LSS to choose the most appropriate methods, which will vary on according to factors such as peptide or protein size, glycosylation state and the theoretical C-terminal amino acid sequence.
C-terminal sequence analysis is a key part of the ICH Q6B guidelines for characterization and confirmation of biopharmaceuticals in support of new marketing applications.

METHODS
LC-MS/MS
HR LC-MS/MS

via Edman degradation
N-Terminal (amino-terminal) sequencing by automated Edman chemistry, also known as automated Gas Phase Sequencing (GPS), is used in protein characterization to determine the amino-terminus of the peptide or protein. It is used throughout all stages of drug discovery or to demonstrate comparability and consistency between batches for release during manufacturing. It is also used during discovery for de novo sequencing of new novel proteins and is available as part of SGS Life Science Services’ full protein characterization package and for antibody analysis.
N-Terminal sequencing can be undertaken on samples in solution or from SDS-PAGE separated proteins blotted on to PVDF membranes. It can be used for the analysis of possible truncations of the amino-terminus, as well as to assess the level of chemically blocked termini which generally prevents full sequencing. It may be possible to remove the blocking group prior to sequencing.
Automated Edman chemistry sequentially breaks down a peptide/protein into its constituent amino acids from the N-terminus of the sample. The produced amino acid is derivatized and separated by RP-HPLC and visualized by UV detection for each cycle of Edman chemistry. The amino acids are quantified by comparison to a standard mixture.
N-Terminal sequencing is a key part of the ICH Q6B guidelines for characterization and confirmation of biopharmaceuticals in support of new marketing applications.

Amino Acid Analysis (AAA) by RP-HPLC is used in protein characterization to analyze peptide or protein concentration, content and molar ratio or, alongside UV spectrophotometry, to gain an extinction coefficient. This analysis is used throughout all stages of drug discovery, as well as to demonstrate comparability and consistency between batches for release during the manufacturing phase.
Amino acid analysis is a necessary part of protein characterization and can also be used for:
• Estimation of extinction coefficient
• Determination of extinction coefficient; the method is validated for the product and may include molecular weight measurement. The validated method may be used for batch release.
• Total amino acids
• Free amino acids
• Protein concentration may be calculated by relating extinction coefficient to Optical Density measurement, which is performed using a UV spectrometer at 280nm. We are also able to scan over a time course and range, 190 - 900nm.
Amino acid analysis is a key part of the ICH Q6B guidelines for characterization and confirmation of biopharmaceuticals in support of new marketing applications.

Peptide mapping by mass spectrometry (peptide mass fingerprinting) is used in protein characterization to produce a unique ‘fingerprint’ of an individual protein and to compare this with the theoretical gene-derived amino acid sequence. This analysis is used for identification purposes at all stages of drug discovery or to demonstrate comparability and consistency between batches for release during manufacturing. It may also be used for the characterization of reference batches.
Peptide mapping is a very powerful tool in protein characterization. Peptide mapping can also be used for:
• Dilsulfide bridge assignment
• N-terminal and C-terminal sequence confirmation (normally together with MS/MS sequencing)
• Screening and identification of post-translational modification sites (e.g. glycosylation, phosphorylation)
• Screening and identification of possible sequence variants and/or truncation/modification sites (e.g. deamidation, oxidation, glycation)
• Guiding the choice of signals for subsequent MS/MS or gas phase sequence analysis
MS Peptide Mapping was originally developed at SGS (through their acquisition of the M-Scan group) in the early 1980’s using Fast Atom Bombardment MS (FAB-MS); the protein molecule is fragmented using specific enzymatic or chemical methods and the resulting peptide mixture is analyzed using MS. Now, the modern technique of Electrospray (ES-MS) is coupled to liquid chromatography (LC-MS).
Peptide mapping is a key part of the ICH Q6B guidelines for characterization and confirmation of biopharmaceuticals in support of new marketing applications.

METHODS

HPLC-MS
UPLC-MS

Protein and peptide de novo sequencing
LC-MS, LC-MS/MS, LC-MSn

Qualitative mass spectrometry

Through our acquisition of the M-Scan group, SGS Life Science Services has unrivalled expertise in protein analysis. Our Philadelphia laboratory was part of a group of originators who developed MS Mapping of biotechnology products, together with other mass spectrometry strategies related to protein/glycoprotein analysis, which have become standards worldwide.
SGS Life Science Services’ protein analysis package is designed to encompass the requirements of the ICH guidelines (particularly ICH Q6B), the US FDA 'Points to Consider' documents and the concept of a 'well characterized' or 'specified' biological product. Individual analyses, or the full protein analysis package, are available to GMP and soon to GLP.

Identification of process and product related impurities, including degradation products (e.g. deamidation, oxidation and mis-matched S-S bridge forms), may be included in the protein analysis package.

SUB-SERVICES
Primary structure
Protein and peptide sequencing
Protein and peptide de novo sequencing
Peptide mapping
Amino acid composition
N-terminal sequencing via Edman degradation
C-terminal sequencing
Disulfide bridge analysis / free thiols measurement
Oligonucleotide analysis
Carbohydrate analysis and PTMs
Monosaccharide composition
Sialic acid analysis
Phosphorylated/sulfated monosaccharide analysis
N- and O-Oligosaccharide qualitative profiling
N- and O-Oligosaccharide quantitative profiling
Linkage analysis
Glycosylation site analysis
Post-translational modifications analyses (qualitative and quantitative)
Physico chemical properties
Molecular weight or size
Isoform pattern
Extinction coefficient estimation or validation
Liquid chromatographic pattern
HOS
Secondary/tertiary structure
Aggregation

Primary structure
Protein and peptide sequencing: Qualitative mass spectrometry
Protein and peptide de novo sequencing: LC-MS, LC-MS/MS, LC-MSn
Peptide mapping: HPLC-MS, UPLC-MS
Amino acid composition: Amino acid analysis
N-terminal sequencing via Edman degradation: Edman degradation
C-terminal sequencing: HR LC-MS/MS
Disulfide bridge analysis / free thiols measurement: LC-MS, LC-MS/MS / Ellman, Fluorescence
Oligonucleotide analysis: LC-MS, LC-MS/MS / ESI-MS, ESI-MS, MALDI-TOF MS

Carbohydrate analysis and PTMs
Monosaccharide composition: HPAEC-PAD, CE-LIF, HPLC-FLD, GC-MS
Sialic acid analysis: HPAEC-PAD, CE-LIF, HPLC-FLD
Phosphorylated/sulfated monosaccharide analysis: HPAEC-PAD, CE-LIF, HPLC-FLD
N- and O-Oligosaccharide qualitative profiling: MALDI-TOF MS
N- and O-Oligosaccharide quantitative profiling: LC-MS, LC-FLD MS, CE-LIF, HPAEC-PAD
Linkage analysis: GC-MS, Enzymatic strategies
Glycosylation site analysis
Post-translational modifications analyses (qualitative and quantitative): LC-MS, LC-MS/MS, LC-MSn, cIEF, iCE, IEX, HIC, RP-HPLC, SEC-HPLC, SDS-PAGE

Physico chemical properties
Molecular weight or size: ESI-MS, LC-MS
Isoform pattern: cIEF, iCE or IEX
Extinction coefficient estimation or validation: AAA
Liquid chromatographic pattern: HPLC/UPLC DAD, UV

HOS
Secondary/tertiary structure: FTIR, CD, Fluorescence
Aggregation: SEC-MALS, SEC-UV, SV-AUC, DLS

Immunogenicity Testing
At SGS Life Sciences, we understand that assessing the immunological responses associated with biological therapeutic products is critical as the responses can ultimately impact the safety and efficacy of a product. Biologics can induce immune responses resulting in the production of anti-drug antibodies (ADA). ADAs can cause unwanted immune responses inactivating the therapeutic effects of the treatment or have adverse effects. In addition to ADAs, evaluation of Neutralizing Antibodies (NAbs) is forms an essential element of immunogenicity testing. NAbs inhibit the biological activity of the biological therapeutic. Thus, during the development of a therapeutic protein, including biosimilars, oligonucleotides, peptides and other biologics, immunogenicity testing needs to be considered at all stages of the drug development pipeline – from preclinical to clinical. Immunogenicity testing of drug products during early and late stage development to commercialization is also recommended by the regulatory agencies.

Our scientists work closely with our clients providing expert guidance to support their immunogenicity analysis requirements throughout the large molecule (biopharmaceutical) development process – from pre-clinical to clinical phases.

ADA Services and expertise include:
• Immunogenicity Assay Development and Optimization
• Immunogenicity Method Transfer
• Immunogenicity Validation
• Routine Analysis
• Multi-tiered approach to measure ADA
• Screening Assay
o Detection of anti-drug antibody (ADA)
o Cut points
• Confirmatory Assay
o Demonstration of ADA specificity
o Use of confirmatory cut points
• Titer or Isotyping
o Determining strength of binding
• Neutralizing Antibody Assay
o Isotyping
o Cross reactivity with endogenous proteins
• Type of Samples: Pre-clinical and clinical sample analysis
• Matrices include rodents and humans
• Platforms: Direct ELISAs and bridging assays, Electrochemiluminescence (ECL) - Meso Scale Discovery (MSD ®)
The main types of assay platforms used in the Immunogenicity testing are ELISA (Bridging Immunoassay and Direct Immunoassay), and Surface Plasmon Resonance. Bridging Immunoassay is used to detect all isotype antibodies, not species-specific and high throughput. Direct Immunoassay is used for high through-put and allows for the detection of low affinity antibodies. Surface Plasma Resonance is used to detect low affinity antibodies. For antibody characterization, cell based Nab assay (direct or indirect) is preferred due to its physiological relevance and ability to detect clinically relevant Nabs. On the other hand, Non-cell based assays (direct or indirect) can be used if cell based assay is not feasible.

Method development, transfer, qualification, validation
Sequence Variants: LC-MS/MS, LC-MSn
Post-translational modifications analyses (qualitative and quantitative): LC-MS, LC-MS/MS, LC-MSn, cIEF, iCE, IEX, HIC, RP-HPLC, SEC-HPLC, SDS-PAGE
Isoform pattern analysis
Glycoform pattern analysis

Quantitative analysis
Residual DNA: ELISA, qPCR
Residual HCP: ELISA, Mass spectrometry
Residual Prot A: ELISA
Residual detergent: HPLC-CAD, HPLC-MS/MS, HPLC-ELSD, HPLC-UV, HPLC-RI, IC, ICP-MS, ICP-OES, GC-MS
Residual antibiotics: HPLC-CAD, HPLC-MS/MS, HPLC-ELSD, HPLC-UV, HPLC-RI, IC, ICP-MS, ICP-OES, GC-MS
Residual process additives: HPLC-CAD, HPLC-MS/MS, HPLC-ELSD, HPLC-UV, HPLC-RI, IC, ICP-MS, ICP-OES, GC-MS
Residual dyes: HPLC-CAD, HPLC-MS/MS, HPLC-ELSD, HPLC-UV, HPLC-RI, IC, ICP-MS, ICP-OES, GC-MS
Residual antifoams: HPLC-CAD, HPLC-MS/MS, HPLC-ELSD, HPLC-UV, HPLC-RI, IC, ICP-MS, ICP-OES, GC-MS
Residual growth promoter/expresser: HPLC-CAD, HPLC-MS/MS, HPLC-ELSD, HPLC-UV, HPLC-RI, IC, ICP-MS, ICP-OES, GC-MS
Residual solubilization agents: HPLC-CAD, HPLC-MS/MS, HPLC-ELSD, HPLC-UV, HPLC-RI, IC, ICP-MS, ICP-OES, GC-MS
Residual Redox reagents: HPLC-CAD, HPLC-MS/MS, HPLC-ELSD, HPLC-UV, HPLC-RI, IC, ICP-MS, ICP-OES, GC-MS
Residual chromarographic purification agents: HPLC-CAD, HPLC-MS/MS, HPLC-ELSD, HPLC-UV, HPLC-RI, IC, ICP-MS, ICP-OES, GC-MS

Qualitative analysis
HCP characterization/identification: Proteomics
Characterization of impurities: LC-MS, LC-MS/MS, LC-MSn, High Resolution MS

Bacterial Endotoxin Testing (BET)/Limulus Amebocyte Lysate (LAL) Test

Throughout the development process and prior to final release to the market, bacterial endotoxin testing (BET) is essential. SGS tests for endotoxin in several areas of drug manufacturing and production including raw material testing, bulk lot release testing and final product release testing for a variety of pharmaceuticals, biologics and medical devices.
Bacterial endotoxin is a fever producing byproduct of Gram-negative bacteria commonly known as pyrogen; detection of endotoxin in samples may be an indicator of the presence of bacterial contaminants within a manufacturing process. It is important to detect any possible contamination during the early stages in time to remediate the situation and take any necessary actions to avoid proliferation. This method replaces the rabbit pyrogen test.
We provide sensitive qualitative and quantitative methods to meet all pharmacopoeial requirements, such as gel clot, kinetic chromogenic and kinetic turbidimetric. With our experienced staff, SGS will be there with you in all phases of drug development to help ensure that your products are pyrogen-free. BET/LAL) testing is conducted in accordance to USP <85>.

Please contact us for guidance regarding sampling and sending samples.

Methods

• Kinetic Turbidimetric
• This technique quantifies the level of bacterial endotoxin present in samples by measuring the increase in turbidity in the presence of a reactant, lysate. A microplate reader is utilized to run the assay.
  o Kinetic Chromogenic
• This technique quantifies the level of bacterial endotoxin present in samples by measuring color change. When endotoxin present in sample reacts with lysate, chromophore is cleaved and causes a yellow color that is measured spectrophotometrically. A microplate reader is utilized to run the assay.
• Gel clot method Gel Clot Method
• This method measures the presence or absence of endotoxins in samples by the development of an opaque gel in the presence of sufficient levels of endotoxin. In cases where a result cannot be obtained using a photometric method, the gel clot method will be used.

At SGS, we can conduct potency assays can be conducted using ligand-binding assays (LBA) or functional assays.

With four GLP laboratories across Europe and USA, SGS are able to serve pharmaceutical and biopharmaceutical companies of all sizes with a range of tests for drug development at both preclinical and clinical stages – from early to late phase. We also develop assays (including immunassays) from scratch.

SGS bioanalysis testing is underpinned by a large list of validated methods and biomarkers, and to maintain our reputation as an industry pioneer we have actively pursued the assay development and validation of some of the more innovative of these biomarkers with our clinical teams. To date we have over 700 assays validated. Our services span:

• Cell-based bioassays
• Immunogenicity testing
• Metabolite profiling
• ADME 14C
• Method transfer, development, optimization and validation
• PD bioanalysis (biomarkers)
• PK bioanalysis (small molecules & biopharmaceuticals)

SGS is the world’s leading inspection, verification, testing and certification company. Our Life Science Services work with the top 20 pharmaceutical companies in the world, but every client receives the same high standard of service – no matter what their size.Pharma and biotech companies of all sizes have chosen us as the preferred bioanalytical partner. SGS has the expertise to both develop assays de novo (including immunoassays) and to support large scale routine sample analyses, from pre-clinical to clinical studies.

In order to support the bioanalysis of various classes of compounds (including peptides), SGS offers a large range of techniques and methods providing rapid high volume bioanalysis. Our method development processes focus on high-throughput technological capabilities (e.g. TurboFlow extraction, multiprobe robots).

SGS offers more than 700 validated bioanalysis methods that are ready for use with a very short lead-time. Validation criteria follow guidelines from the FDA (May 2001) and the Crystal City Conference (May 2006). All validations are tailored to specific program requirements.

Pharmacokinetic (PK) and Toxicokinetic (TK) studies are usually conducted at the discovery or drug candidate selection stages of the drug development lifecycle. PK and TK studies provide insight into fate of a drug upon administration.

In order to support PK and TK studies of biopharmaceuticals, a wide range of immunochemistry platforms such as ELISA, EIA, ECL, Luminex, LC-MS and methods combined with scientific expertise are available. Services include:
o Pre-clinical and clinical support
o Custom Immunoassay Method Development
o Method Transfer
o Method Optimization
o Method Validation - Validation criteria follow guidelines from the FDA and the Crystal City Conference. All validations are tailored to specific program requirements.
o Routine Testing
o SGS offers validated bioanalysis methods that are ready for use

Dried Blood Spot Analysis
Dry Blood Spot (DBS) technology has several advantages for toxicokinetic (TK) and pharmacokinetic (PK) analysis, including: reduced blood sampling collection, absence of post-collection processing, low biohazard risk, fewer required facilities for sample shipment and storage. Furthermore, sample analysis can also be simplified.

Early to Late Phase Clinical Trials and Post-Marketing

To support the bioanalysis of various classes of compounds, a large range of techniques and methods are available to you. Also, an exceptional sample processing and analytical capabilities, combined with scientific expertise, enables us to provide rapid, high-volume bioanalysis. Each service is overseen by highly skilled and trained skilled analysts, who are able to offer rapid high volume analyses that use a range of innovative techniques, including:

• Mass Spectrometry
• LC-MS/MS
• GC-MS
• Immunoanalysis
• Immunochemistry
• Ligand Binding Assays
• Singleplexed Custom Immunoassay Development
• EIA’s
• Radioimmunoassay (RIA)
• Biomarker assay development and validation
• Multiplexed Custom Immunoassays
• BD Beads array and quantification by BD FACSCanto II + High Throughput Sampler for 96- and 384-well
  microtiter plate capacity driven by BD FACSDIVA software, fully validated according to 21 CFR Part 11 rules
• Biomarker assay development and validation
• Classical HPLC
• HPLC systems equipped with UV, fluorescence and electrochemical detectors
• UPLC
• Automated sample preparation using robotics (multiprobe) or turbulent flow technology (cohesive)
• Large sample storage capacity combined with excellent sample tracking (400,000 sample storage and equivalent backup)
• Current throughput of more than 200,000 incurred samples per year
• Sample handling and tracking by the Watson LIMS

SGS Life Sciences offers bioanalytical support for every stage of large molecule drug development process - early discovery through phase I, II, III and IV to the pharmaceutical and biopharmaceutical companies of all sizes.

Our analytical platforms include ELISA, MSD-Electrochemiluminescence and multiplexing capabilities and focus on bioanalytical studies for monoclonal antibodies, proteins, conjugated proteins and peptides.

Our GLP-compliant Bioanalytcal Immunochemistry Laboratory services include:
• Pharmacokinetic (PK) bioanalysis
• Toxicokinetic (TK) studies
• Immunogenecity testing
• PD bioanalysis (biomarkers)
• Method transfer
• Method development
• Method optimization
• Method validation
• Routine Testing
• Quantitative and Qualitative immunoassays
• GLP and non-GLP Immunoassays to support clinical and pre-clinical studies
Our laboratories use Watson LIMS for sample handling and tracking.

In vivo Toxicity Testing Services

Toxicology Services

Product Development & Testing Services

Product Testing Services

Protein Purification and Quantification Services

Pharmacology Services

Drug Discovery & Development Services

Manufacturing Services