Assay developers invest millions of dollars perfecting their analytical methodologies, optimizing primer sequences, refining detection algorithms, and validating performance under well controlled conditions. Despite this investment, many find that their assay, which performed well in the lab, exhibits unexpectedly variable results in clinical settings. In our experience, the root cause often lies not with the assay itself but with an often-overlooked aspect of the diagnostic workflow – pre-analytics.
The Pre-Analytical Challenge: Where Perfect Assays Meet Reality
“Pre-analytics” refers the activities and conditions occurring prior to the actual analytical testing of a sample. In assay development, pre-analytics research entails investigating variables that may affect assay performance such as collection method (e.g., tube type), labeling system, transport (including type of container), processing (at specimen site or in the testing laboratory), and storage conditions. In the controlled environment of a research laboratory, these elements are standardized and optimized; biospecimens are quickly processed using tightly drafted SOPs and stored under well-controlled conditions. In clinical practice, however, the situation can be quite different.
Real-world clinical deployment sites — hospital laboratories, testing facilities, physician offices, and other sites — have their own individualized workflows, capabilities, and operating constraints. A hospital lab processing hundreds of samples every day operates differently than a research facility evaluating a handful of specimens a day. Staffing, equipment, training, and mindset vary radically and the ensuing sample variability can impact biospecimen integrity, biomarker stability, and ultimately, assay performance. This is why what worked in the lab may fail in the “real world.’
Critical Pre-Analytical Variables in Biospecimen Management
Our experience helping our customers cross the bridge between the laboratory and the clinic has elucidated several key pre-analytical variables that can impact performance, including:
- Biospecimen Collection and Processing Windows
The journey from biospecimen collection to processing is more variable than developers anticipate. In controlled research settings, samples are processed immediately or at least within a couple hours. In the clinical world, however, biospecimens often sit for much longer periods. This delay can have significant effects on biomarker stability. Circulating tumor DNA can degrade, cell-free DNA concentrations can change due to ongoing cell lysis, and protein biomarkers may become unstable. The challenge for the developer is determining how much delay the new assay can tolerate while maintaining performance.
- Biospecimen Shipping and Transportation Conditions
Biospecimens often need to travel—sometimes across the country—to reach the testing laboratory. During this journey, samples may be exposed to temperature fluctuations, vibration, and rough handling. Room temperature shipping may be cost-effective and simple, but is it compatible with your biomarker’s stability? Refrigerated transport offers better temperature control but increases complexity and costs.
- Biospecimen Processing Protocol Variations
Variations in biospecimen processing can also impact results. Clinical sites do not all use the same centrifuge models and regularly spin at different speeds or spin times. Some laboratories have equipment for automated sample handling while others work with manual set ups. These variations can dramatically affect biospecimen quality and composition, altering biomarker concentrations or introducing artifacts that impact assay performance.
- Biospecimen Storage Protocol Compliance and Monitoring
Biospecimen storage represents another point where variability can creep in. Temperature fluctuations during storage, freeze-thaw cycles due to equipment issues, and storage durations all have the potential to compromise biospecimen integrity.
- Biospecimen Collection Tube Selection and Anticoagulant Effects
The choice of collection tubes and anticoagulants also affects biomarker stability and measurement. EDTA tubes may be preferred for DNA-based assays while heparin tubes might be better for protein biomarkers. Some anticoagulants can interfere with downstream processes or affect the stability of biomarkers during biospecimen storage. Some tubes like those from Streck or PreAnalytiX offer longer stability profiles but come at a higher price. Clinical sites may also have preferences or constraints that limit their choice of collection tubes, making it important to understand how these variations might impact your assay’s performance.
The MT Group Approach: Expert Biospecimen Management
Recognizing the importance of pre-analytical variables, MT Group has developed studies to evaluate these factors’ impact assay performance. Our approach goes beyond standard analytical validation and explores the entire biospecimen journey from collection to result. The net effect is minimization of Garbage In, Garbage Out (GIGO), a common outcome when commercialization-enabling assay demonstrations are built upon sub-optimal pre-analytics.
Controlled Comparative Biospecimen Studies
We conduct A vs. B collection studies that allow the direct comparison of different specimen pre-analytical variables. By collecting and handling biospecimens from the same patients but under different conditions — immediate vs. delayed processing, different storage temperatures, collection tube a vs. b — our sponsors can quantify the impact of each variable on assay performance.
These comparative biospecimen studies are essential for the development and validation of novel assays. They provide the data needed to establish standard operating procedures, define processing windows, and identify which pre-analytical factors are most important for real-world deployment. Our studies also generate data about biospecimen handling tolerances, which is valuable for developing clinical protocols, training clinical site personnel, and establishing quality control procedures that ensure performance across deployment sites.
The Business Case for Pre-Analytical Validation
While conducting pre-analytical validation studies requires time and resources, the investment pays dividends:
- Reduced Clinical Validation Failures: By identifying and addressing pre-analytical issues before clinical deployment, companies can avoid failures and performance inconsistencies that could damage their reputation and delay broad market adoption.
- Broader Market Access: Assays that are robust to pre-analytical variations can be deployed more widely, such as in clinical settings with limited biospecimen handling capabilities. This flexibility can expand the potential market.
- Regulatory Advantages: Pre-analytical validation data strengthens regulatory submissions and demonstrates an understanding of real-world conditions.
- Customer Confidence: Clinical laboratories and healthcare providers are more likely to adopt assays that come with evidence-based pre-analytical protocols and demonstrated robustness.
Looking Forward: The Evolving Biospecimen Landscape
Assays are being deployed in diverse clinical settings, from academic medical centers to community hospitals to point-of-care testing environments. Each setting presents challenges for biospecimen collection, processing, and management and success in this evolving landscape requires an understanding of how pre-analytical variables impact assay performance and a commitment to developing real-world-ready test protocols. Companies that invest in pre-analytical validation studies will be better positioned to achieve clinical success and capture market opportunities.
Partnering for Success
At MT Group, we understand that developing and validating assays to assist clinical practice is a challenge. Our team brings decades of experience in bridging the gap between laboratory research and clinical implementation, helping companies navigate the pre-analytical considerations and biospecimen management strategies that can determine the success or failure of their diagnostic products. By partnering with specialists who understand both the requirements of assays and the realities of clinical biospecimens, assay developers can avoid pitfalls and accelerate their path to market.
Success in the market depends not only on assay excellence, but also on understanding and management of the entire diagnostic workflow — from the moment blood is drawn to the delivery of results. Companies that recognize and address this reality will be the ones that transform patient care through their diagnostic solutions.
For more information about MT Group’s biospecimen collection capabilities to support pre-analytical development, contact our team of specialists.
