Good Clinical Laboratory Practice Research Articles

TitrationAnalysis: a tool for high throughput binding kinetics data analysis for multiple label-free platforms.

Label-free techniques including Surface Plasmon Resonance (SPR) and Biolayer Interferometry (BLI) are biophysical tools widely used to collect binding kinetics data of bimolecular interactions. To efficiently analyze SPR and BLI binding kinetics data, we have built a new high throughput analysis tool named the TitrationAnalysis. It can be used as a package in the Mathematica scripting environment and ultilize the non-linear curve-fitting module of Mathematica for its core function. This tool can fit the binding time course data and estimate association and dissociation rate constants ( k a and k d respectively) for determining apparent dissociation constant ( K D) values. The high throughput fitting process is automatic, requires minimal knowledge on Mathematica scripting and can be applied to data from multiple label-free platforms. We demonstrate that the TitrationAnalysis is optimal to analyze antibody-antigen binding data acquired on Biacore T200 (SPR), Carterra LSA (SPR imaging) and ForteBio Octet Red384 (BLI) platforms. The k a, k d and K D values derived using TitrationAnalysis very closely matched the results from the commercial analysis software provided specifically for these instruments. Additionally, the TitrationAnalysis tool generates user-directed customizable results output that can be readily used in downstream Data Quality Control associated with Good Clinical Laboratory Practice operations. With the versatility in source of data input source and options of analysis result output, the TitrationAnalysis high throughput analysis tool offers investigators a powerful alternative in biomolecular interaction characterization.

Candor in medical laboratory management “Quality is doing the right thing and making things right”

The practice of medicine in the modern era is exclusively evidence based, focuses on justifiable laboratory reports contributing effective and opportune patient management. For many years, clinical laboratories have picked up astounding significance in medical care services. Laboratory investigations are essential for medical diagnosis in patient care as well as medical research. Reliability of laboratory data/report is therefore of paramount importance. Good Clinical Laboratory Practices (GCLP) is essential to be implemented in the field of medical research and health care services to ensure reliability of laboratory data. Medical Laboratory Professionals endorse the decency, decorum, honour, morality, and self-respect of the profession and maintain a integrity, competence, reputation of honesty, authenticity, and accuracy. All laboratories engaged in testing of biological samples need to establish confidence in the quality and reliability of the results of these tests. In the current time, ethical concerns exist everywhere whether it is a medical field or life science. Medicine, at the center of which is the sick and dying man, has developed humanism and the morality of pity for the afflicted "Noli noceri",(First do no harm)"Salus aegroti - supreme lex"( let the welfare of the people be the supreme law ). It coincides with the motto of the World Academy of Medical Sciences mission. "Dedication to duty in a mission for mankind".

Effect of Good Clinical Laboratory Practices Quality Training on Knowledge, Attitude and Practice among Laboratory ProfessionalsQuasi Experimental Study

Introduction: Good Clinical Laboratory Practices (GCLP) play a vital role in early and accurate diagnosis, providing highquality data, and timely sample processing. Adhering to a robust Quality Management System (QMS) that complies with GCLP standards is crucial for laboratory personnel in a clinical laboratory to deliver outstanding healthcare services and reliable, reproducible reports. Aim: To assess the Knowledge, Attitude, and Practice (KAP) of laboratory professionals towards quality in the laboratory through GCLP training. Materials and Methods: This pre-test, post-test quasiexperimental study was conducted in the Department of Biochemistry at ESIC Medical College and Hospital, Faridabad, Haryana, India, from February 2022 to June 2022. The study included 58 participants, consisting of 22 doctors and the remaining laboratory assistants. GCLP online training program was conducted every Friday in March 2022 for four weeks. An online questionnaire containing 34 questions was administered to all the participants before and after the training. Data were collected and analysed using a paired t-test. Results: A total of 58 responses were received from the participants via Google form before and after the training. The results indicate no significant difference in participants’ responses to 12 closed-ended questions regarding QMS before and after training. A similar trend was observed for 22 questions on a Likert scale, where participants rated their agreement, neutrality, or disagreement. Conclusion: The study demonstrates that all technical staff fully complied with GCLP guidelines and accreditation requirements. Furthermore, the laboratory staff acknowledges the importance of Standard Operating Procedures (SOPs), document maintenance, record-keeping, and identifying nonconformities, all of which contribute to effective traceability of the testing process in the clinical laboratory.

The design and implementation of natural population cohort study Biobank: A multiple-center project cooperation with medical consortia in Southwest China.

The West China Hospital of Sichuan University collaborated with regional medical consortia in Sichuan Province to launch a natural population cohort study (NPCS) to investigate the health status of residents and collect public health data in southwest China. Up to 80,000 participants will be enrolled by the NPCS from 11 regional medical consortia over five years. Individuals are invited to visit one of 11 participating medical consortia to fill out questionnaires, receive a free health exam, and donate biospecimens upon enrolment. All participating medical facilities adhered to standard operating procedures for collecting and processing biospecimens to ensure uniformity (serum, lithium heparinized plasma, ethylene diamine tetraacetie acid plasma, and buffy coat). The Electronic Data Capture System, Picture Archiving and Communication System, Laboratory Information Management System, Biospecimen Quality Control System, Biobank Information Management System, and will be used to sort and classify clinical indices, imaging data, laboratory parameters, pre-analytical variables, and biospecimen information, respectively. All quality assurance and quality control procedures in the NPCS biobank adhered to the "DAIDS Guidelines for Good Clinical Laboratory Practice Standards". This project will integrate high-dimensional multi-omics data, laboratory data, clinical data, questionnaire data, and environmental risk factors. An estimated 2,240,000 aliquots of the sample will be stored by the end of the study. These samples are linked with comprehensively collected clinical indices, imaging data, and laboratory parameters. Big data analysis can be implemented to create predictive algorithms, explore pathogenesis mechanisms, uncover potential biomarkers, and provide information on public health. NPCS will provide an integrative approach to research risk factors and pathogenesis of major chronic or endemic diseases in Sichuan Province and provide key scientific evidence to support the formulation of health management policies in China.

A Comparative Review of ICMR, WHO, and EMA Guidelines for Good Clinical Laboratory Practices.

With the advancement of clinical research and the increased burden on laboratory services, there is an unmet need for guidelines regarding proper laboratory functioning and reliable data generation. Several organizations from all over the world have published guidelines for these clinical and research laboratories. Good Clinical Laboratory Practices (GCLP) are stepwise procedures aimed at strengthening the quality of test results produced by all clinical laboratories engaged in human sample analysis. In this article, we attempt a comparison of the GCLP guidelines recently issued by the Indian Council of Medical Research with the guidelines released by the World Health Organization and the European Medicines Agency. Also, we have included and discussed several suggestions that, if included, will lead to the strengthening of the laboratory practices used for both research and patient care for overall improvement in the Indian healthcare system.

Achieving intracellular cytokine staining assay concordance on two continents to assess HIV vaccine-induced T-cell responses.

The HIV Vaccine Trials Network (HVTN) conducts clinical trials on 4 continents in pursuit of a safe and effective HIV vaccine. Cellular immune responses to vaccination that define vaccine immunogenicity and/or immune correlates of protection can be measured using multiparameter intracellular cytokine staining (ICS) assays. The HVTN cellular immunology laboratory, located in Seattle, WA, conducts ICS assays for vaccine trials according to Good Clinical Laboratory Practices (GCLP). In 2013, the HVTN established a second GCLP compliant cellular immunology laboratory in Cape Town, South Africa to assess vaccine immunogenicity for HVTN trials conducted on the African continent. To ensure ICS readouts in the 2 laboratories were directly comparable, we conducted concordance testing using PBMC from healthy controls and vaccine trial participants. Despite standardized procedures and instrumentation, shared quality control measures and quality assurance oversight, several factors impacted our ability to obtain close agreement in T-cell responses measured in the 2 laboratories. One of these was the type of fetal bovine serum (FBS) used in the assay, which impacted lymphocyte cell viability and background responses. In addition, the differences in supernatant removal technique also significantly affected our ability to detect positive responses to vaccine antigens. Standardization of these factors allowed us to achieve and maintain ICS assay concordance across the 2 laboratories over multiple years, accelerating our efforts to evaluate HIV vaccines. The insights gained in this process are valuable for assay transfer efforts by groups of investigators that need to directly compare data generated in different laboratories around the globe.

Capacity Building in Sub-Saharan Africa as Part of the INTENSE-TBM Project During the COVID-19 Pandemic

Tuberculous meningitis (TBM) is the most severe and disabling form of tuberculosis (TB), with at least 100,000 cases per year and a mortality rate of up to 50% in individuals co-infected with human immunodeficiency virus type 1 (HIV-1). To evaluate the efficacy and safety of an intensified anti-tubercular regimen and an anti-inflammatory treatment, the INTENSE-TBM project includes a phase III randomised clinical trial (TBM-RCT) in four countries in sub-Saharan Africa (SSA). Within this framework, we designed a comprehensive capacity-building work package ensuring all centres had, or would acquire, the ability to conduct the TBM-RCT and developing a network of skilled researchers, clinical centres and microbiology laboratories. Here, we describe these activities, identify strengths/challenges and share tools adaptable to other projects, particularly in low- and lower-middle income countries with heterogeneous settings and during the coronavirus disease 2019 (COVID-19) pandemic. Despite major challenges, TBM-RCT initiation was achieved in all sites, promoting enhanced local healthcare systems and encouraging further clinical research in SSA. In terms of certified trainings, the achievement levels were 95% (124/131) for good clinical practice, 91% (39/43) for good clinical laboratory practice and 91% (48/53) for infection prevention and control. Platform-based research, developed as part of capacity-building activities for specific projects, may be a valuable tool in fighting future infectious diseases and in developing high-level research in Africa.Supplementary InformationThe online version contains supplementary material available at 10.1007/s40121-022-00667-z.

Comprehensive Development and Implementation of Good Laboratory Practice for NGS Based Targeted Panel on Solid Tumor FFPE Tissues in Diagnostics.

The speed, accuracy, and increasing affordability of next-generation sequencing (NGS) have revolutionized the advent of precision medicine. To date, standardized validation criteria for diagnostic accreditation do not exist due to variability across the multitude of NGS platforms and within NGS processes. In molecular diagnostics, it is necessary to ensure that the primary material of the FFPE sample has good quality and optimum quantity for the analysis, otherwise the laborious and expensive NGS test may result in unreliable information. Therefore, stringent quality control of DNA and RNA before, during, and after library preparation is an essential parameter. Considering the various challenges with the FFPE samples, we aimed to set a benchmark in QC metrics that can be utilized by molecular diagnostic laboratories for successful library preparation and high-quality NGS data output. In total, 144 DNA and 103 RNA samples of various cancer types with a maximum storage of 2 years were processed for 52 gene focus panels. During the making of DNA and RNA libraries, extensive QC check parameters were imposed at different checkpoints. The decision tree approach can be set as a benchmark for FFPE samples and as a guide to establishing a good clinical laboratory practice for targeted NGS panels.

Strengthening laboratory capacity for HIV vaccine clinical trials and epidemiological studies in Eastern and Southern Africa

Background: Conducting successful HIV vaccine clinical trials in resource-limited settings is hampered by lack of adequate laboratory capacity at trial sites, poor infrastructure, lack of well-trained technical personnel, and inadequate laboratory quality management Systems. We describe our approach to establishing sustainable laboratory capacity for clinical trials in Africa. Methods: IAVI identified 9 CRCs where a capacity building program that supports immunology and clinical testing was established. Information from the 9 CRCs was collected retrospectively and compiled in Microsoft excel for descriptive statistics. Mapping was done in Quantum Geographic information system. Results: Newly built and refurbished laboratories have been equipped with the required testing laboratory equipment. All CRC laboratories (n=10, 100%) received Good Clinical Laboratory Practice (GCLP) accreditation between 2004 and 2016, and accreditation maintained annually. A total of 89 audits were done between 2005-2019. KAVI and KEMRI had the highest number of audits (n=11, 12.4%). IAVI successfully trained a total of 1811 individual, of which (n=1130, 62.7%) trained on GCLP, (n=330, 18.3%) Quality Management Systems, (n=311, 17,2%) laboratory techniques and (n=32,1.8%) between 2004 and 2021. All the 13 Assays were registered in either College of American pathologist (CAP) or Royal college of pathologists of Australasia (RCPA) for Proficiency testing. Conclusion: The establishment of GCLP accredited laboratories and well-trained personnel has created centers of excellence and it has enabled them to attract independent competitive research funding. The GCLP accreditation and standardized testing procedures ensured reliable and accurate data, especially important for multi-country and multi-center studies.

Preparing for Clinical Laboratory Improvement Amendments (CLIA) and College of American Pathologists (CAP) Inspections.

Clinical laboratories have many regulations to follow, requiring complete adherence to specific standards and regulations in order to be granted accreditation. As part of the accreditation process, a laboratory must be inspected. Whether it is an initial or biennial inspection, there are some standard tasks and duties a laboratory can do to prepare in advance to reduce stress, improve the inspection process, and reduce the risk of getting a deficiency. Good Clinical Laboratory Practice (GCLP) is an important part of preparing a clinical laboratory for Clinical Laboratory Improvement Amendments (CLIA) and College of American Pathologists (CAP) inspections. GCLP standards have been developed by CLIA and were developed with the goal of providing a sole source of requirements that clinical laboratories using human patient samples need to follow to ensure reproducible and reliable clinical laboratory results. The Laboratory Accreditation Program (LAP) of the College of American Pathologists (CAP) also has ongoing activities and guidelines for clinical laboratories to follow. Although this is a voluntary program, it is driven by peer review, education, and compliance to established performance standards. CAP is focused on laboratory improvement and views its inspections as collaborations between inspector and laboratory. The CAP checklists, based on their standards for good lab practice, are used by inspectors to ensure that each inspection is consistent and thorough and to enable CAP to determine if the laboratory meets the standards for accreditation. © 2021 Wiley Periodicals LLC.

The impact of online management systems: a qualitative assessment of staff perception at a clinical research laboratory

Good Clinical Laboratory Practices (GCLP) increase the quality and traceability of results in clinical research. However, high turnover of staff, insufficient resources, and lack of training in lab management may limit its implementation at Academic Health Centers (AHCs). This work aimed to qualitatively assess the staff perception of the implementation of a freeware Online Management System (OMS) on the workflow of an academic clinical research laboratory. A free online OMS (Quartzy, Quartzy Inc., USA) was selected and implemented from 2012-2016. After training interventions, a qualitative analysis was performed for the staff attitude towards the implementation, including a structured questionnaire (30 participants) and focus group assessments (16 participants). Indicators of management performance were also compared before and after the implementation. The results indicate that lab members perceive improvement in organization, communication, and commitment of staff, who reported the system as user-friendly and a facilitator for the autonomy of group members. On the other hand, there is a recognition of the need for constant training and engagement at the management level. These findings suggest that full-scope OMS may represent useful tools to assist in staff engagement in compliance with GCLP at AHC laboratories.

A Circular Network of Coregulated Sphingolipids Dictates Lung Cancer Growth and Progression

Sphingolipid metabolism is one of the top dysregulated pathways in NSCLC. However, the molecular control of sphingolipid metabolic reprogramming in cancer progression remains unknown. Here, we first provide clinical evidence that expression of sphingolipid metabolic enzymes, B3GNT5 and GAL3ST1, but not the others, is most significantly associated with patient prognosis, whilst sphingolipidomics analysis of NSCLC patient sera identifies their metabolites, neolacto-series glycosphingolipid and sulfatide, as biomarkers that outperformed current clinical biomarkers for staging patients. Further network analysis of the sphingolipidomes reveals a circular network of coregulated sphingolipids, indicating that the neolacto-series glycosphingolipid/sulfatide balance functions as a checkpoint to determine sphingolipid metabolic reprograming during patient progression. Importantly, sphingolipidomics analysis of B3GNT5/GAL3ST1 genetically perturbed NSCLC cell lines confirms their key regulatory role in sphingolipid metabolism, while B3GNT5 and GAL3ST1 expression has an opposite role on tumorigenesis in vitro and in vivo. Overall, our results provide new insights whereby B3GNT5 and GAL3ST1 differentially regulate sphingolipid metabolism in lung cancer growth and progression. Funding Statement: This work was supported by the Natural Science Foundation of China (81920108028, 81872142); Guangzhou Science and Technology Program (201904020008); Young Teacher Foundation of Sun Yat-sen University (18ykzd07); Guangdong Science and Technology Department (2020B1212060018, 2019A1515011802, 2020A1515011280, 2020A0505100029); China Postdoctoral Science Foundation (2019M650226, 2019M650227). Declaration of Interests: No potential conflicts of interest were disclosed. Ethics Approval Statement: Serum samples were collected from healthy individuals and NSCLC patients (age-matched and body-mass index –matched) with informed consent, and the research program and all the related procedures were approved by the Ethics Committee of Sun Yat-sen Memorial Hospital (Guangzhou, China) and stored according to standard operating procedures for Good Clinical Laboratory Practice standards. All animal procedures were approved by the Institutional Animal Care and Use Committee (IACUC) of Sun Yat-Sen University.

Biobanking for Translational Diabetes Research in India

India is declared as the diabetic capital of the world. Clinically well-annotated blood samples will advance diabetes research for better diagnostic and treatment methods. Building a disease-specific biobank with high-quality peripheral blood mononuclear cells (PBMCs) and clinical follow-up data system will serve as a good platform for clinical research in diabetes. Processing and storage of high-quality biospecimen for translational research in diabetes demand the implementation of good clinical laboratory practices. “Certification or accreditation programs” that improve biorepository processes and frameworks are lacking in Indian context. To sustain and translate the research into clinical practice, good governance of the biobank and financial resources is required. For ethical issues related to health needs of the people and participants in the research, issues related to research process, translational research, and commercialization, data sharing should be addressed. For India to be an innovation and sustainable country Indian government is supporting translational research facilities, including biobanks. India has developed biobanks for various diseases; however, diabetes-specific research biorepository is lacking. Given the dangers of diabetic burden, India should set up a diabetes disease-specific repository learning from the global organizations and customize to the needs of Indian context. It is important to have private agencies get involved to develop biobanks and future research as there are commercial goals to translate research into practice. New technologies of specimen storing and preservation, data management, and data sharing should be adopted for developing cost-effective long-standing disease-specific population biobank in India.

International Council for Standardization in Haematology (ICSH) recommendations for laboratory measurement of ADAMTS13.

This guidance document was prepared on behalf of the International Council for Standardization in Haematology (ICSH), by the ADAMTS13 Assay Working Group, which comprises an international group of both clinical and laboratory experts. The document provides recommendations on best practice for the performance of ADAMTS13 assays in clinical laboratories. ADAMTS13 assays support the differential diagnosis of thrombotic microangiopathies and have utility in the management of thrombotic thrombocytopenic purpura (TTP). There are three types of assay: activity, antigen and autoantibody/inhibitor assays. Methods for activity assays differ in terms of sensitivity, specificity, precision and turnaround time. The most widely used assays involve VWF peptide substrates and either chromogenic ELISA or FRET techniques, although chemiluminescence assays and rapid screening tests have recently become available. Tests for autoantibodies and inhibitors allow confirmation of acquired, immune-mediated TTP, while antigen assays may be useful in congenital TTP and as prognostic markers. In this document, we have attempted to describe ADAMTS13 assays and the conditions that affect them, as well as: blood collection, sample processing, quality control, standardization and clinical utility; recognizing that laboratories in different parts of the world have varying levels of sophistication. The recommendations are based on expert opinion, published literature and good clinical laboratory practice.

Diagnostic tests for COVID-19: the importance of the before and the after

In 2019, in the city of Wuhan, China, patients were diagnosed with a novel coronavirus originally named 2019-nCoV, currently known as SARS-CoV-2. The alarming expansion of this pandemic makes it necessary to find out and implement reliable diagnostic methods in order to properly detect and treat patients, thus contributing to slowing down the spread of the disease. It is extremely important to have trained professionals in sample collection, good clinical laboratory practices, and molecular and hematological techniques to adequately detect any cases of infection with this virus. Moreover, standardized protocols are essential for obtaining, transferring and storing samples to stop the spread of the COVID-19 pandemic caused by SARS-CoV-2.

Pre-positioned Outbreak Research: The Joint Medical Emerging Diseases Intervention Clinical Capability Experience in Uganda.

The West Africa Ebola virus disease outbreak of 2014-2016 demonstrated that responses to viral hemorrhagic fever epidemics must go beyond emergency stopgap measures and should incorporate high-quality medical care and clinical research. Optimal patient management is essential to improving outcomes, and it must be implemented regardless of geographical location or patient socioeconomic status. Coupling clinical research with improved care has a significant added benefit: Improved data quality and management can guide the development of more effective supportive care algorithms and can support regulatory approvals of investigational medical countermeasures (MCMs), which can alter the cycle of emergency response to reemerging pathogens. However, executing clinical research during outbreaks of high-consequence pathogens is complicated and comes with ethical and research regulatory challenges. Aggressive care and excellent quality control must be balanced by the requirements of an appropriate infection prevention and control posture for healthcare workers and by overcoming the resource limitations inherent in many outbreak settings. The Joint Mobile Emerging Disease Intervention Clinical Capability was established in 2015 to develop a high-quality clinical trial capability in Uganda to support rigorous evaluation of MCMs targeting high-consequence pathogens like Ebola virus. This capability assembles clinicians, laboratorians, clinical researchers, logisticians, and regulatory professionals trained in infection prevention and control and in good clinical and good clinical laboratory practices. The resulting team is prepared to provide high-quality medical care and clinical research during high-consequence outbreaks.

Implementation of a three-tiered approach to identify and characterize anti-drug antibodies raised against HIV-specific broadly neutralizing antibodies

The ability to detect, quantify, and interrogate the properties of immune responses raised against biological therapeutics is not only important to our understanding of these molecules, but also to their success in the clinic. A tiered assay approach to identify the presence, specificity, and titer of anti-drug antibody (ADA) responses has been adopted as a gold standard by industry leaders, the FDA, and the EMA. In order to support pre-clinical and clinical trials, these assays must be standardized, and their performance sufficiently characterized to ensure the accuracy and reproducibility of results under relevant testing conditions. Here we present implementation of electrochemiluminiscence assays that fit into the tiered paradigm of ADA testing for five HIV broadly neutralizing antibodies (3BNC117, 3BNC117-LS, 10–1074, PGT121, and PGDM1400) in compliance with Good Clinical Laboratory practices. Assay sensitivities and matrix effects were evaluated and used to inform the development of positivity cut points. Once cut points were established, assay precision, specificity, free-drug tolerance, and robustness were defined. In all cases, assay characteristics met or surpassed recommendations set forth by the FDA. To further evaluate the performance of these assays and the tiered approach, samples from non-human primates that had received a subset of the five therapeutics were evaluated. In sum, this study reports qualification of a set of ADA assays available to the scientific community as pre-clinical and clinical trials of broadly HIV-neutralizing antibodies proceed, and a framework that is easily adapted as new drug products are advanced in the clinic.

Optimization and qualification of a functional anti-drug antibody assay for HIV-1 bnAbs

The recent identification of human monoclonal antibodies with broad and potent neutralizing activity against HIV-1 (bnAbs) has resulted in substantial efforts to develop these molecules for clinical use in the prevention and treatment of HIV-1 infection. As with any protein therapeutic drug product, it is imperative to have qualified assays that can accurately detect and quantify anti-drug antibodies (ADA) that may develop in patients receiving passive administration of HIV-1 bnAbs. Here, we have optimized and qualified a functional assay to assess the potential of ADA to inhibit the neutralizing function of HIV-1 bnAbs. Using a modified version of the validated TZM-bl HIV-1 neutralization assay, murine anti-idiotype antibodies were utilized to optimize and evaluate parameters of linearity, range, limit of detection, specificity, and precision for measuring inhibitory ADA activity against multiple HIV-1 bnAbs that are in clinical development. We further demonstrate the utility of this assay for detecting naturally occurring ADA responses in non-human primates receiving passive administration of human bnAbs. This functional assay format complements binding-antibody ADA strategies being developed for HIV-1 bnAbs, and when utilized together, will support a multi-tiered approach for ADA testing that is compliant with Good Clinical Laboratory Practice (GCLP) procedures and FDA guidance.

The mapping procedure as an essential process during clinical trials

A risk–based approach to the quality of clinical trial (CT) data provides the appropriate conditions for the use of materials at clinical trial site (CTS). One of the most important parameters requires careful monitoring of the temperature mode for storing samples. In case of any temperature failure there can be the data loss that may affect quality of any CT. The procedure of temperature mapping enables to confirm the uniformity and stability parameters of the temperature distribution within the freezers at CTS.Aim. To analyze the guidelines regulating the quality of CT and develop a standard operating procedure (SOP) of the mapping process for freezing equipment at the CTS.Materials and methods. At the Clinical and Diagnostic Center (CDC) of the NUPh the temperature mapping was performed according to the requirements of the WHO in order to prevent risks associated with temperature fluctuations in storage areas of biological samples. This procedure is quite important for the further installation of portable temperature registration systems (data loggers), which are necessary to monitor the temperature regime during CT.Results. At the moment, the documents regulating the mapping procedure are relevant for pharmaceutical manufacturing. However, continuous scientific and technical development, as well as automation of equipment requires the development and implementation of the mapping process into the CT process. Temperature mapping is carried out through a number of requirements, which are partially mentioned in various guidelines on the quality of medical products and the operation of freezing equipment at CTS – guidelines of good practices: Good Clinical Laboratory Practice (GCLP), Good Manufacturing Practice (GMP), Good Storage Practice (GSP), Good Distribution Practice (GDP) and Good Automated Manufacturing Practice (GAMP). The latest 2012th edition of the International Society for Pharmaceutical Engineering (ISPE) GAMP–5 is a set of guidelines for manufacturers and users of automated systems in the pharmaceutical industry. All of these good practices establish the mapping procedure as a required part of the operational and performance qualifications of the freezing equipment used to store drugs in manufacturing, storage facilities and during their transportation, particularly as a part of the “cold chain”. As far as properly calibrated freezing equipment is used during the CT, mapping can also be considered as part of the qualification in a regulatory procedure at CTS. We have developed SOP, which determines the requirements and the algorithm for conducting the mapping procedure at CTS.Conclusions. The current requirements of ISPE GAMP–5 pharmaceutical engineering indicate that mapping is an automated process that has several advantages over the manual data capturing of temperature fluctuations. The mapping process allows continuously recording and storing temperature values at a specified interval for several days using temperature sensors, as well as it helps: to assess the stability of freezers; to identify critical (cold and hot) zones with the largest deviations of the temperature from the acceptable operating range; to determine critical (control) points for the purpose of further installation of data loggers to control the maintenance of a given temperature regimen of freezers. Thus, temperature mapping reduces the risk of errors, eliminates the “human factor” during the temperature recording in the freezer, as well as mobilizes the work of CTS. Further studies will make a background of a single national mapping document that is necessary nowadays to minimize the risks of any participant of CT.

Customisation and validation of a low-volume plasma renin activity immunoassay: Enabling of regulatory compliant determination in paediatric trials

ObjectivesInvestigations of plasma renin activity (PRA) in children are urgently required. Small-volume, regulatory guideline compliant, bioanalytical assays tailored for paediatric application could facilitate to overcome this hurdle. Ethical constraints given e.g. by the European Medicines Agency need to be addressed and reliable data generation in line with Good Clinical Laboratory Practice must be ensured. MethodsA PRA enzyme-linked immunosorbent assay (ELISA) was tailored for paediatric application and validated in the context of the U.S. Food and Drug Administration bioanalytical guideline. Performance verification of the assay was conducted by participation in an interlaboratory ring test, evaluation of incurred sample reanalysis and an application-orientated approach in children. ResultsA five-fold reduction of required plasma volume to 100 μL was achieved without limiting the calibration range. Between-run accuracy and precision varied no more than 5.0% and 6.3%, respectively. No substantial matrix effect was detected and the inter-run precision for parallelism was 11.1%. Stability experiments approved the freeze-thaw stability, short-term stability as well as 37 weeks of long-term stability. The assay successfully participated in the interlaboratory ring test, showing non-inferiority regarding radioimmunoassay (RIA). Moreover, PRA in plasma samples of neonates was successfully determined. Conducted incurred sample reanalysis confirmed the comparability and reliability of the assay with regard to international regulatory bioanalytical guidelines. ConclusionA fit-for-purpose PRA ELISA characterised by low-volume application was successfully established, indicating non-inferiority regarding commonly applied RIAs. Reliability of the regulatory-compliant PRA assay was proven by participation in an interlaboratory ring test and its application in a paediatric population.