Case Studies
We have a proven track record in assay development for diverse clinical studies, including gene therapy, respiratory and orphan drugs, immunogenicity testing, dermatology, arthrosis trials, and prostate cancer screening.
Case Study 1
Patient-Centric Microsampling
Bioanalysis (2022) 14(20), 1305-1315 https://www.future-science.com/doi/abs/10.4155/bio-2022-0167
Collection of PK samples outside of the clinical setting was required to support the rapid development of Paxlovid for the treatment of Covid-19 in multiple populations. Unilabs' scaleable prioritisation, adaptability and resilience offered validation of a new technique and bioanalysis in record time for an accelerated submission schedule.
Approach
- Device Selection: Tasso M-20 device chosen for its ease-of-use in sample collection. It uses tiny needles to penetrate the skin and microfluidics to generate a slight vacuum that pulls blood from capillaries into the cartridge.
- Sample Collection: Blood is deposited onto 4 pads containing volumetric absorptive micro sampling material (VAMS) to collect dried blood samples (~20 µL).
- Method Development: Specific spiking techniques were developed for preparing calibration and QC samples.
- Validation Assessments: Evaluations for hematocrit effect and sample stability were conducted at various conditions (−80°C, −20°C, ambient 10–30°C, elevated 40°C, and 90% RH humidity).
- Bridging Studies: Studies established concordance between dried blood and plasma PK data.
Outcome
- Initiated in late Q1 2021 and validated to current regulatory standards.
- Bridging studies confirmed a high degree of concordance between dried blood and plasma.
- Applied in multiple clinical studies with PK collection performed by patients or caregivers at home.
- Paxlovid received FDA Emergency Use Authorisation in December 2021 and full approval in May 2023.
Case Study 2
Peptide Drug Conjugates
Overview
This study covers the progression of peptide drug conjugates from discovery through to clinical evaluation, using different bioanalytical strategies.
Approach
Metabolism/Disposition Studies in Drug Discovery
- In Vivo Characterisation: Conducted metabolite characterisation studies in animals to assess metabolic liability/disposition.
- Non-Regulated Bioanalysis: Established payload exposure in relevant animal tissues.
- Definitive Binding Studies: Employed equilibrium dialysis to characterise binding for both the conjugate and the payload.
Quantitative Bioanalysis and Biotransformation for Clinical Studies
- Assay Development: Developed and validated sensitive LC-MS/MS assays for the intact conjugate and payload.
- Exposure Determination: Measured conjugate and payload exposure in clinical samples to inform safety and efficacy.
- Metabolite Characterisation: Performed follow-up studies on drug-related material in clinical samples.
Case Study 3
Peptides
Overview
This study focuses on metabolism/disposition studies for peptides and peptide–drug conjugates through in vivo investigations and biotransformation studies.
Approach
Vivo metabolite characterisation studies
- Biotransformation Studies with peptide Containing Drugs: In vivo Case Study with a Bicyclic Toxin Conjugate. Mark Savage: DMDG Biotransformation Workshop, 2021
- Mass Spectrometry Processing Software for Peptide Metabolite Identification. Jan Boerma: Peptide ADME Discussion Group, 2021
Protein Binding Determinations
- Performed using equilibrium dialysis and ultracentrifugation
Case Study 4
Increasing Drug Tolerance for ADA Studies
Overview
Addressing the development of two novel cancer therapies and detection of anti-drug antibody concentrations in samples impeded by high concentrations of drug ("drug tolerance"). This study focused on developing drug tolerant ADA assays because of the long half-life of the therapeutics, which increased drug levels in samples.
Approach
Case Study 1:
- Increased drug tolerance by using acid dissociation.
- Increased the minimum required dilution (MRD) and augmented the concentration of labelled drug in the Master Mix.
Case Study 2:
- Employed Solid Phase Extraction with Acid Dissociation (SPEAD).
- Samples underwent initial acid dissociation followed by capture with biotinylated drug and streptavidin-coated beads, plus a further acidification step.
Outcome
Case Study 1:
- Achieved drug tolerance of 1000 µg/mL.
Case Study 2:
- Achieved drug tolerance of 120 µg/mL.
- Both approaches were successfully validated with the necessary drug tolerance achieved.
Case Study 5
First-in-Class Bispecific Fusion Protein for Melanoma
Overview
A bispecific fusion protein targeting a melanoma-associated antigen employs a dual mechanism: a T-cell receptor (TCR) domain along with an anti-CD3 T-cell engaging domain, redirecting T cells to target tumour cells expressing the melanoma-associated antigen. Unilabs agility, responsiveness and tailored, customer-centric approach supported the registration journey in lock-step with our client.
Approach
Method Development:
- Initially developed and validated in mouse serum in 2010.
- Early preclinical PK studies were supported by this method.
Clinical Application:
- Subsequently, the method was developed and validated in human serum.
Supported numerous clinical trials by providing fast turnaround PK data.
Outcome
The drug received approval from both the FDA and EU in 2022 for the treatment of metastatic uveal melanoma, and it is currently undergoing trials for treatment of other cancers.
Case Study 6
Biomarker antibody levels impacting reproducibility in hybrid assays
Overview
This case study focuses on troubleshooting an LC-MS/MS method transferred for a protein biomarker in human plasma. The method initially performed well but encountered issues after transferring due to matrix-specific challenges.
Approach
Initial Assessment:
- Method was transferred using dog plasma for calibration and human plasma (pre-screened for low endogenous biomarker levels) for QCs.
- During validation, all bulk spiked QC levels were below the LLOQ.
Troubleshooting Steps:
- No matrix effects seen in prior method evaluation or Sponsor's validation
- Re-spiked QCs in same matrix; all QCs remained below LLOQ
- Identified immunocapture stage as source of biomarker loss
- Prepared QCs in different control matrix; all QCs within limits
- Tested QCs in ten matrices (including suspect); all calculated correctly except suspect matrix (below LLOQ).
- Diluted suspect matrix with control plasma up to 100-fold and spiked QCs; levels increased above LLOQ but remained significantly reduced
- Literature Review: Biomarker antibodies found in patients with undiagnosed disease state (1% prevalence); antibody levels greatly reduced with life-long treatment.
Outcome
The method was subsequently validated to current guidelines.
No further instances of suspect control matrices have been observed, although the effect remains theoretically linked to an undiagnosed patient subpopulation.
Case Study 7
CDx Testing to Reintroduce MS Drug
Overview
When Tysabri® was withdrawn due to JC-virus risk, Unilabs worked with Biogen to implement wide-scale CDx testing. This solution enabled Tysabri®'s return to market and continues to manage risk in multiple sclerosis patients through global anti-JCV testing.
Approach
Method Transfer & Validation:
- December 2010: Unilabs was selected following Biogen’s RFP.
- February 2011: Transfer and validation of the anti-JCV risk test method.
Full-Service Operational Model:
- Encompasses collection material, sample logistics, analytical results, and longitudinal monitoring.
Global Coverage & Rollout:
- March 2011: Initial launch with 280 EU physicians and parallel lab service during IVD approval.
- May 2011 onward: Expanded to 8,000+ physicians across 70+ countries with under 7-day turnaround times.
Data Analytics:
- In-market dynamics reporting provides strategic insights to pharmaceutical companies.
Outcome
Over 50,000 patients receive annual safety testing.
More than 1,5 million Stratify JCV tests have been performed, enabling Tysabri®’s global reintroduction and sustaining Unilabs’ competitive advantage through its tailored web portal for Health Care Providers.
Case Study 8
CDx Testing to Reintroduce MS Drug
Stratify Testing Process
Unilabs Stratify website:
A customised web solution for HCPs to sign up for the Stratify JCV & JCV DNA test, request sampling material, register patient samples online, schedule sample pick-up, and view test results.