Research biobanks are public (non-profit) or private institutions that gather, handle, store, analyze, and disseminate systematically organized biological specimens and data for use in research and development projects. The main aim of biobanks over “traditional” sample collections is to ensure the integrity of both biological and associated clinical data, thus supporting a variety of research areas by providing high-quality samples, consistent data, and dedicated expertise. This scientific scope is achieved in the context of a solid quality system and in respect of the participants’ rights and ad hoc regulations (1). Indeed, the biological material and related data are processed solely by authorized and adequately trained personnel; access to data storage systems and privileges are controlled by appropriate security measures and total traceability of the sample from collection to storage, is guaranteed by uniform procedures such as best practices (2) and International Organization for Standardization (ISO) standards (namely ISO 9001:2015 and UNI 20387:2018).

Research biobanks usually deal with specific diseases or restricted cohorts of donors/patients (3, 4); the sample stored following biobanking standards ensure reproducibility of scientific results and can be transferred to other researchers (following strict rules to ensure donors privacy) and assembled with other sources to build up large cohorts, in order to meet broad research needs (extending beyond those of the hosting institution) spanning from elucidation of pathogenetic mechanisms, to development and validation of innovative diagnostic tools, discovery of biomarkers for prognostic and therapeutic stratification, and development of novel therapeutics (5). Other advantages include the involvement of stakeholders, patients’ associations (especially regarding ethical and legal aspects), and potential commercial partners. Finally, biobanks represent the basis for the development of national and international networks since standardized processes and procedures are easier to share among structures. Actually, in 2013, biobanks were proposed to be networked by the European Commission with a pan-European project, the Biobanking and BioMolecular Resource Research Infrastructure- European Research Infrastructure Consortium (BBMRI-ERIC) for a transition from individual research tools to complex international research infrastructures. However, this is a complex and challenging process and is still at an early stage (6).

As a result, biobanks are a strategic resource for influencing the long-term path of medical research and a powerful substrate for high throughput OMICS studies and integration with rapidly improving big data handling techniques, including artificial intelligence and deep le (lines)arning methods, to reach the goal of “personalized medicine” for an ever-growing number of human diseases (7). The strategic nature of biobanks is indeed recognized by the European Community in its framework programs encouraging their development in terms of quality, interoperability, and livelihood, with the most recent example in the ninth framework program Horizon Europe 2021–2027 (pillar “Excellent Science”) focused on research quality or the Italian National Recovery and Resilience Plan (PNRR) (8).

On this premises, we established a rheumatology BioBank, led by the Medicine and Rheumatology Unit of the IRCCS Istituto Ortopedico Rizzoli (IOR) in Bologna, Italy (RheumaBank@IOR), with the goal of collecting, processing, storing, and distributing biological samples and associated data obtained from patients with rheumatic diseases. The RheumaBank, along with two other biobanks (Genetic Biobank and Musculoskeletal Tumor Biobank), is part of the coordinating unit Biological Resources Centre (CRB), member of BBMRI and affiliated to a pilot project promoted by the Italian Ministry of Health (MOH) for the accreditation to the UNI 20387:2018 “Biotecnology-Biobanking-General requirement for Biobanking” standard (henceforth referred to as UNI 20387). This participation is the first step toward obtaining UNI 20387 certification, which is critical for recognizing biobank’s operating quality standards (9). Whole blood, serum, urine and isolated peripheral blood mononuclear cells (PBMC) are currently collected in the RheumaBank, after obtaining specific informed consent from patients. RheumaBank donors are patients suffering from rheumatoid arthritis (RA), psoriatic arthritis (PsA) and other axial or peripheral spondyloarthritis (SpA), that cumulatively account for the vast majority of patients with inflammatory joint diseases seen in rheumatology clinics worldwide.

To present, the RheumaBank has already enrolled over 800 donors and stores over 4,200 samples divided into about 14 aliquots for each sample. These numbers include historical collections related to prior research projects carried out at the Medicine and Rheumatology unit, as well as samples newly enrolled in the Biobank from 2022 onwards. Samples are stored in special −80°C ultra freezers and in liquid nitrogen containers located in restricted rooms with limited access. The samples collected and processed by the Rheumatology Biobank are available to the scientific community for use in rheumatology research projects, and their traceability and controlled storage characteristics ensure research with reliable and repeatable results, a prerequisite for the transferability of knowledge to clinical practice.

The aim of this paper is to describe the methodologies used to establish the path for collection, handling, and storage of biological samples in RheumaBank, in accordance with quality standards that ensure traceability and monitoring throughout the process.

In 2022, a total of 246 patients were enrolled in the RheumaBank. Four different sample types were collected from each patient: whole blood (2 aliquots), serum (7 aliquots), urine (3 aliquots) and PBMC (2 aliquots) for a total of 3444 aliquots at the T0 timepoint, as represented in Table 4. Recruitment for T3, T6, and T12 is ongoing. The number of patients enrolled for each rheumatic disease reflects the prevalence of the condition in the general population, with RA having the highest prevalence (Table 4).

More than 80% of serum, blood and urine samples were processed within 4 h from collection, and PBMC samples were prepared and stored within 48 h from blood collection in about 90% of samples.

The quality tests performed on serum samples showed a coefficient of variation (CV) under the standard medium CV (20%) as shown in Figure 2.

A better understanding of human diseases is strictly dependent on depicting the pathophysiological processes and recognizing the relationship between genotype and phenotype, which, in turn, could lead to the successful development of novel medications and more effective personalization of available treatments. Biobanks play a critical role in ensuring sampling homogeneity and reliability through the implementation of best practices, as well as providing access to a large number of biological samples, thereby accelerating research and allowing for the rapid translation of results into clinical practice (41). In this context, the United Kingdom (UK) biobank is such an excellent example, collecting genotype of about 500,000 individuals associated with extensive phenotype (lifestyle, biomarkers, imaging) data. The UK biobank has already promoted the collection of multiple data sets on the relationship between phenotype and pathogenic genotype, significantly improving our understanding of human functioning in health and disease, confirming the enormous value of this research infrastructure (42).

Rheumatic diseases are common in the general population, yet despite the availability of novel therapeutic options, a significant proportion of patients continue to fail to respond effectively to first-line treatments, pointing to one of the field’s biggest unmet needs (43). Given the heterogeneity of arthritis pathogenesis, the goal of therapeutic strategies should ideally be to target the pathogenic mechanism prevailing in each individual patient. Further, some individuals may have diverse processes concurring to the disease phenotype, resulting in a partial or complete response to a narrowly targeted therapy (44). These factors have resulted in an urgent need for a better understanding of the pathophysiology of these diseases, as well as the identification of biomarkers that can reliably predict the best treatment for a given patient and provide a theoretical basis for the development of novel therapeutics.

In this manuscript we describe the main activities carried at our Institution, a specialistic musculoskeletal research hospital, to build a rheumatology biobank. To the best of our knowledge, RheumaBank is the first biobank in Italy specifically conceived to provide support to basic, clinical, and translational studies in rheumatology, aiming at shade light on the pathophysiological mechanisms and stimulating the development of novel therapeutic molecules in a personalized medicine framework. However, to set up a biobank is a complex task, that requires taking in account several aspects, from those concerning governance and ethics to pure methodological, operational, and technical ones (45).

The first important point to consider is to build an informed consent form taking into consideration all ethical, legal, and social issues while adhering to national and international standards. In our case, we prepared this document in compliance with the guidelines of the Ethical, Legal and Social Issues (ELSI) working group of BBMRI.it, the national node of BBMRI-ERIC, the European research infrastructure for biobanking and biomolecular resources (46). All the preparatory information necessary to sign the consent is provided to patients by healthcare professionals skilled in the field of biobanking; additionally, to encourage patient compliance, blood sampling is scheduled to harmonize with routine health care appointments (47).

Recent articles have emphasized the importance of the reproducibility of scientific data, which is closely linked to the quality of the biological samples (48). Published studies have been discovered to be difficult or impossible to replicate or reproduce, a methodological crisis known as the “replication crisis” (also known as the replicability crisis or reproducibility crisis) (49). Even though the outcomes of preclinical research cannot be replicated, many secondary papers are nonetheless created (50). To counteract this phenomenon, research biobanks use a predetermined procedure for collecting, handling, and storing biological samples that ensures the standardization of the methods, specific training of personnel, and continuous checks on the path and the samples (51, 52). Furthermore, biobanks implement quality controls (on both banked samples and collected data) throughout banking procedures to verify the quality of the samples and, consequently, the reliability of scientific results and their application (53). While the quality controls that must be carried out on cells and nucleic acid samples are well described in the reference CEN standards (54, 55), the publication of a specific standard for serum and plasma samples is still pending.

Our workflow is designed to guarantee sample processing within 4 h after collection. In most applications, this meets the requirements for research use. In cases where stricter conditions are mandatory, such as serum samples used for metabolomics analysis where processing within 90 min is indicated (56), it will be possible to choose the idoneous samples since detailed processing times are always recorded. In this way, those who will have to use biobank samples for research purposes will always be aware of when they were processed and be able to decide which analyses can or cannot be performed (57). Accordingly, we run ad hoc experiments to evaluate the stability of specific molecules relevant to the diseases collected in the biobank (58).

All the standard operating procedures developed and described here have been periodically evaluated and improved. The whole process complies with the ISO 9001:2015 standards and an upgrade to the UNI 20387 standard is underway. It should be noted, however, that the standardization and improvement of pre-analytical procedures for in vitro studies is an ongoing process. The most up-to-date high priority pre-analytical CEN and ISO standard documents, as well as the corresponding External Quality Assessment (EQA) schemes and implementation tools were the basis for the establishment of our pathway. Not only following adequate standard operating procedures (SOPs) is essential to guarantee research results, but also having qualified personnel, aware of the role and potential of the biobank (59, 60). To reach this goal, a continuous training plan has been foreseen. In particular, the operative biobank personnel attended several training courses on UNI 20387, organized by Accredia, the sole national accreditation body appointed by the Italian government in compliance with the application of the European Regulation 765/2008, and allowed to certificate the compliance to UNI 20387. Furthermore, every member of the RheumaBank staff obtained the certificate to conduct internal audits for the assessment of compliance with the UNI 20387 standard.

Finally, the working environment is undeniably one of the most important aspects of biobank success, requiring the strict collaboration of all involved personnel, beginning with medical staff charged with selecting donors and collecting fundamental clinical data; to healthcare personnel charged with acquiring informed consent, to nursing staff charged with collecting biological samples, to research and technical personnel charged with sample manipulation and storage. The complex biobank workflow will succeed only if all of these components work together to achieve the same aim.

The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.

The studies involving humans were approved by Comitato Etico di Area Vasta Emilia Centro della Regione Emilia-Romagna (CE-AVEC). The studies were conducted in accordance with the local legislation and institutional requirements. The participants provided their written informed consent to participate in this study.

EA, SNa, FU, and SNe: conceptualization and methodology. EA, SNa, and SNe: investigation and data curation. FU: resources and funding acquisition. EA and SNe: writing—original draft preparation. EA, SNa, VB, JC, LL, LM, FP, LP, CF, FU, and SNe: writing, review, and editing. EA, SNe, and FU: project administration. All author has been involved either in the conception and design of the study, or in the acquisition of data, or in the analysis and interpretation of data, or in drafting the article or in revising it critically for important intellectual content, or in the final approval of the submitted version.

This study was funded by the IRCCS Istituto Ortopedico Rizzoli, Bologna–Italy (“Ricerca corrente” fund).

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

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