Biospecimen standardization has emerged as a critical priority in clinical trials, with the implementation of

ICH E6(R3) guidelines in 2025 marking a paradigm shift toward enhanced data integrity and quality

management. This comprehensive review examines current best practices, regulatory frameworks, and

from collection through analysis, while addressing practical considerations for multi-site clinical trials. By

establishing harmonized practices, the clinical research community can enhance data reliability, facilitate

global collaboration, and ultimately improve the translation of research findings into clinical practice.

Export, Quality Management, Digital Biobanking, International Regulations, Sample Integrity

Biospecimen standardization in clinical trials refers to the systematic implementation of harmonized protocols,

procedures, and quality standards across all phases of the biospecimen lifecycle—from collection and

processing through storage, distribution, and analysis. This encompasses the establishment of evidence-based

standards, and chain-of-custody protocols. The field has experienced rapid evolution, with the 2025

implementation of ICH E6(R3) guidelines representing a watershed moment that emphasizes quality-by-design

principles and comprehensive data governance throughout the clinical trial lifecycle.

The regulatory environment governing clinical trials and biospecimen management has undergone substantial

transformation with the adoption of ICH E6(R3) Good Clinical Practice guidelines on January 6, 2025. This

revision introduces innovative provisions designed to apply across various types and settings of clinical trials,

ensuring continued relevance amid ongoing technological and methodological advancements.

ICH E6(R3) emphasizes a Quality-by-Design approach, requiring sponsors to proactively identify and mitigate

risks throughout the trial lifecycle, moving beyond traditional prescriptive models toward flexible, risk-based

Pre-analytical variables represent the collection, processing, and storage conditions that occur before

laboratory analysis and constitute the most significant source of variability in biospecimen research. The

National Cancer Institute's Biospecimen Pre-analytical Variables (BPV) Program has systematically

investigated these factors through rigorously controlled studies, examining the effects of cold ischemic time

(delay to fixation), time in formalin, freezing methods, and storage temperatures on molecular profiles.

The BPV Program established a robust biospecimen collection infrastructure to prospectively collect

specimens using rigorous standard operating procedures, controlling for most variables while introducing

experimental conditions to study specific handling issues. This program has analyzed DNA and RNA from

biospecimens collected under controlled conditions using multiple molecular platforms, with findings

the first comprehensive series of procedural guidelines annotated with published findings in human

biospecimen science. These guidelines organize literature findings by specific pre-analytical factors and

analytes of interest—including DNA, RNA, protein, and morphology—providing detailed, adaptable formats

intended to support the development and execution of evidence-based standard operating procedures.

Research demonstrates that quantitative reverse transcription-PCR assays show superior sensitivity in

assessing RNA quality, consistently detecting differences between FFPE (formalin-fixed paraffin-embedded)

and snap-frozen biospecimens across various time points. Traditional quality metrics like RNA Integrity

Number and DV200 (representing the percentage of RNA fragments longer than 200 nucleotides) display more

limited sensitivity, highlighting the importance of selecting appropriate quality assessment tools for specific

applications.

are classified by priority: required fields (Priority 1), clinically important but not yet validated (Priority 2), not

Fig 2.1: Standardized Collection Workflow

tube type, processing delay, and storage conditions all demonstrating measurable effects on analytical results.

For liquid biopsies utilizing cell-free DNA, preanalytical considerations are particularly challenging, as

inconsistency between sample handling protocols and lack of standardization among analytical techniques

that a 12-hour delay to fixation can adversely affect molecular quality in specific tumor types, while optimal

fixation times must be established for each tissue type and analytical platform. Snap-frozen tissue requires

immediate immersion in liquid nitrogen or storage on dry ice, with documentation of the freezing method

essential for quality assessment.

protocols, with some metabolites showing substantial instability without preservatives. The use of borate or

chlorhexidine preservatives can significantly alter metabolite profiles, necessitating careful protocol selection

based on intended downstream analyses.

The International Society for Biological and Environmental Repositories (ISBER) has established the most

Fig 3.0: Comprehensive Laboratory Management Overview

Biorepositories face the substantial challenge of maintaining biospecimen integrity and quality over extended

time periods, often spanning decades. A robust quality management system incorporating both quality

assurance (QA) and quality control (QC) represents the foundation for preserving biospecimen quality

throughout the storage lifecycle.

for all processes, train staff comprehensively on those SOPs, and review documents periodically to ensure

currency and relevance. SOPs serve to standardize biobanking methods and minimize variation in biospecimen

molecular integrity. Frozen biospecimens typically require storage at -80°C or in liquid nitrogen vapor phase (-

196°C), with the choice depending on intended duration of storage and analytical applications. Real-time

The guidelines emphasize that clinical trial processes, measures, and approaches should be implemented

proportionate to risks, avoiding unnecessary burden on participants and investigators. This risk proportionality

ensuring protection of trial participants' data confidentiality, managing computerized systems appropriately,

The guidelines require robust procedures covering the entire data lifecycle—from data capture with

appropriate metadata and audit trails, through data correction and transfer, to eventual finalization for analysis.

For biospecimen operations, this translates to:

The packaging and shipment of biospecimens constitutes a multistep process governed by distinct regulations

depending on whether specimens are shipped domestically or internationally and whether shipments contain

hazardous materials. Failure to comply with these regulations may result in shipment delays, confiscation, or

destruction by quarantine officers at ports of entry, along with potential civil or criminal penalties.