Liquid nitrogen is commonly used in biorepositories for the storage of cells and tissue samples. However, liquid nitrogen handling is associated with risks of serious accidents, including cold burns, asphyxiation, and explosions. These concerns have recently been brought to the limelight by a fatal nitrogen gas leak that occurred at a Georgia poultry plant in January 2021. Strict precautions should be taken while working with liquid nitrogen, including the use of personal protective equipment and compliance with safe handling procedures.

          Liquid nitrogen properties

          Nitrogen is an odorless, colorless, and tasteless gas that is liquified under high-pressure conditions. Liquid nitrogen is liquefied nitrogen gas with a boiling point of approximately -196°C (-320°F). It undergoes spontaneous gas conversion with a rate depending on the specific conditions, which decreases the amount of liquid nitrogen.¹ Notably, liquid nitrogen can expand into large gas volumes.

          Liquid nitrogen has found applications in many areas of human life, including in medicine (for removal of certain skin lesions), as a computer coolant, in cryogenics (the production and effects of very low temperatures), and in the storage of biospecimens.

          Significance of liquid nitrogen in biorepositories

          Liquid nitrogen plays an important role in the storage of both cells and tissue samples in biorepositories. It preserves biospecimens at a very low storage temperature that halts biological activity – a major advantage of liquid nitrogen as a cryopreserving agent. The exact mode of liquid nitrogen handling in biorepositories varies with the scale and type of tissue storage conditions. Facilities with multiple cryofreezers opt for bulk tank storage of the liquid nitrogen outside the storage facilities. Vacuum-jacketed piping will be installed to distribute the liquid nitrogen to individual freezers. Such a set-up is much safer, since the bulk of liquid nitrogen placed outside the enclosed areas of biorepositories. Biorepositores with a limited number of cryofreezers opt for smaller, portable dewar systems to hook up to individual cryofreezer. It is critical to secure the dewars to prevent them from rolling off and causing tipping hazards. In any of these use cases, working with liquid nitrogen is always  associated with clear risks that require strict precautions.

          A recent example of a fatal accident occurred at a poultry plant in Georgia² clearly illustrates the risks associated with liquid nitrogen use and significance of addressing this issue diligently. During unplanned maintenance work on a processing and freezing line, a nitrogen gas leak took place that caused six fatalities and 12 hospitalizations.

Risk of accidents associated with liquid nitrogen use^1,3

          Handling liquid nitrogen is associated with an increased risk of accidents, including asphyxiation, cold burns, and explosions. In case of a liquid nitrogen-related accident, medical attention should be sought immediately.

          Risk of asphyxiation

Liquid nitrogen can expand into large volumes of gas, which can lead to oxygen displacement and risk of asphyxiation.³ Therefore, in cases of large liquid nitrogen spills, the personnel should be evacuated. Moreover, if people working with liquid nitrogen become dizzy, they should be immediately evacuated to a well-ventilated area.

          Risk of cold burns, frost bites, and eye damage

          The very low temperature of liquid nitrogen leads to another important hazard: risk of cold burns, frost bites, or eye damage after contact with liquid nitrogen due to instant freezing.³ If a person’s skin has been exposed to liquid nitrogen or cold nitrogen gas, the normal body temperature should be restored by flashing the affected area with large volumes of tepid water, and the tissue should be protected from further damage and infection. However, the frozen body part should not be rubbed to prevent further tissue damage, and heat should not be applied.

          Risk of explosion due to pressure buildup or due to liquid oxygen

          When liquid nitrogen expands into gas in an enclosed environment, risk of explosion arises. Liquid nitrogen should not be stored in sealed containers because sealed containers may not be able to contain the pressure caused by its gas expansion. A pressure relief vessel or a venting lid should be used to protect against pressure buildup.

Biobanks and natural, technological, and “man-made” disasters
          The biotech and pharmaceutical industries are facing an increased need for high-quality biospecimens for research program success, which is driving the demand for advanced biostorage infrastructure. Access to biospecimens is a limiting factor in many research fields, and biobanks play an important role for the collection, processing, and storage of biospecimens. Moreover, biobanks make biospecimens available to researchers, promoting scientific progress in many pharmaceutical areas.

          Optimal operations of biobanks rely on risk mitigation and emergency planning for different types of crises, including natural (biological, geological, or meteorological); technological (hardware, software, power, or telecommunications); and human derived (intentional or accidental) disasters¹. Biobanks take several steps to design, practice, and implement a well-defined strategies including risk identification, risk assessment, and development of contingency and recovery plans. Accidents and disasters can be difficult to predict, so having a plan for when disruptive or destructive instances occur is a core competency for biorepositories. Biorepositories will learn best from one another, using Lessons Learned from another repository’s real crisis response helps the whole industry create better mitigative best practices and emergency plans for the future.

Risk identification and assessment by biobanks
          Identification and assessment of natural, technological, and human crises risks are performed by each biobank based on geographic location, types of storage offered, and types of samples stored. The probability and the potential impact of risks are taken into consideration in the course of this assessment², and  help crisis management teams streamline and prioritize a rapid and well-coordinated response. All potential risks, including those with low probability or low impact, are addressed with a mitigation strategy and failure plan.

Development of contingency and recovery plans by biobanks
          The development of contingency plans is critical in mitigating risks and aiding recovery efforts^1,3. Implemented strategies include the establishment of detailed standard operating procedures (SOPs), continuous monitoring of equipment, back-up power and equipment, priority lists with emergency contacts, alternative storage solutions, and efficient communication^1–4.

Detailed and updated SOPs
          SOPs are updated to include the findings of a risk assessment and a detailed and tested emergency response plan. Performing routine tests and drills of the emergency plan included in an SOP ensures a well-practiced, effective crisis response.

Continuous monitoring of the equipment
          Continuous monitoring ensures that the storage equipment that are the foundation of biobanking are functioning properly and that any malfunction is detected promptly. When a technical malfunction is identified, the incident is captured on Non-Conformity forms and CAPA is initiated if the issue is severe enough or has an impact on more than one area of the biobank. Continuous monitoring of freezer temperature is an example of a high-severity risk mitigation practice to prevent the loss of valuable biospecimens. A single freezer cooling too much or not cooling enough could severely affect the quality of the specimens inside.

Establishment of back-up systems
          Back-up equipment, including freezers, are always available to ensure that samples can be transferred quickly in the case of a system malfunction. Moreover, emergency generators are used and tested on a regular basis to ensure a smooth transition of power if an outage occurs. Liquid nitrogen and CO₂ may also be provided for the storage units to provide emergency cooling. All back-up systems are checked regularly to ensure their proper function in the case of an emergency.

Provision of back-up supplies
          Back-up supplies are either available or a plan for their provision is in place in cases of emergency. These supplies an additional backup emergency power unit (i.e., a generator) with priority delivery and back-up liquid nitrogen are examples of such supplies.

Cancer is the second leading global cause of death, accounting for approximately one in six deaths worldwide.¹ Biomarkers represent defined characteristics that indicate physiological or pathogenic processes, medical conditions, or responses to interventions or exposures. Biological molecules assessed in tissues or body fluids, including blood, may serve as biomarkers. Notably, biomarkers play an important role in cancer research because they may facilitate the development of novel or optimized therapeutic strategies. Moreover, biomarkers may aid the implementation of precision medicine, which develops prevention and treatment strategies taking into consideration individual variability.² Indeed, due to the high mortality, morbidity, and disability rates associated with cancer, oncology has been the area of most active development of precision medicine and biomarker research.²

Biomarkers in oncology
Many cancer biomarkers provide a powerful and dynamic approach to understand the complexities associated with malignancies and have already been utilized in practice with diagnostic, prognostic, predictive of a treatment response, or monitoring value. For example, human epidermal growth factor receptor 2 (HER2) positivity is predictive of trastuzumab response in patients with breast cancer. Moreover, in colorectal cancer, KRAS-activating mutations are predictive of resistance to treatment with epidermal growth factor receptor (EGFR) inhibitors.³ Another example for effective use of multiple cancer biomarkers is the Oncotype DX gene panel, which is a 21-gene genomic assay predicting the risk of recurrence of breast cancer.⁴

Challenges faced in cancer biomarker research
The development of biomarkers for cancer research is a time- and effort-consuming process. Moreover, the vast majority of potential biomarkers that show promise in early studies fall short of the expectations and cannot be implemented in the clinical practice. The reasons are complex and may include exaggeration of the significance of early findings, overrepresentation of male study participants in clinical trials, and limited access to biospecimens for cancer biomarker research.⁴

How can biobanking aid cancer biomarker research?
Timely and efficient access to biospecimens is essential for the identification of novel biomarkers. Accordingly, professionally procured and well-annotated biospecimens have been shown to contribute to the findings of approximately 40% of all cancer research papers.⁵ Modern biobanks have played a significant role in supporting such activities.

Challenges biobanks face in cancer biomarker research and possible solutions
The organization and operation of biobanks have evolved over the years and have progressed from afterthoughts to a scientific discipline. As this field matures, the complexities may vary according to the biobank type. Some of the challenges biobanks face with regard to cancer biomarker research relate to the type and quality of biospecimens, their availability, and biobank sustainability.

Challenges in the identification of cancer biomarkers and the potential of -omics technologies
The identification of cancer biomarkers has been impeded by the complex pathophysiological pathways implicated in neoplastic processes. However, the progress of high-throughput -omics technologies, including genomic, proteomic, metabolomic, and multiplex approaches, has enabled the screening of a large number of potential targets and facilitates the identification of cancer biomarkers.⁶

Necessity to identify biomarkers in less-invasively or non-invasively collected biospecimens
The collection of biospecimens to assess molecular cancer biomarkers may be invasive and time- and effort-consuming. Liquid biopsy, in which cancer cells or cancer cell-derived DNA fragments are determined in blood, is less invasive.⁷ Thus, the identification of cancer biomarkers in liquid biopsies would reduce the burden on patients and facilitate longitudinal follow up of the biomarker.

Limited access to biological specimens may impede both the diagnosis of many medical conditions and the development of novel therapeutics.¹ Biobanks address this problem by collecting, extensively characterizing, storing, and disseminating biological specimens; thus, facilitating research development. BioArkive, San Diego’s leading biorepository and pre-clinical research service provider that offers cost-effective, end-to-end biobanking solutions.

The rapid spread of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has led to the coronavirus disease 2019 (COVID-19) pandemic, severely affecting both global healthcare and economy.  The COVID-19 pandemic has also posed numerous challenges to biobank operations, necessitating adjustments in their management and workflows. However, biobanks have also contributed to progress in COVID-19 research in addition to the support it was providing for a broader array of medical and disease research. BioArkive has also continued to serve its customers throughout the COVID-19 pandemic and to provide high-quality biobanking, drug discovery, and cold-chain logistics solutions.

The contribution of biobanks to combating the COVID-19 pandemic
Several biobanks have been created to collect specimens for COVID-19 research, in addition to the efforts existing biobanks had already undertaken to respond to COVID-19 related cold-chain logistics and storage needs.⁴ Thus, biobanks play an important role in the collection, characterization, storage, and dissemination of specimens that can be used for COVID-19 research. In the long run, this may aid in the development of diagnostic and therapeutic tools for combating COVID-19.¹

Challenges in the field of biobanking caused by the COVID-19 pandemic
Biosafety concerns

Collecting and processing biological specimens have been associated with significant biosafety concerns during the pandemic. Thus, handling samples from patients with a SARS-CoV-2 infection requires the biobank personnel to follow strict safety guidelines.² The exposure risk for a SARS-CoV-2 infection extends to handling biological specimens from clinically healthy individuals or from individuals with unrelated medical conditions, and it is now best practice to assume that any biological specimens collected, processed, and stored by biobanks may be infected. Moreover, appropriate protocols have been activated or developed to handle potentially or confirmed infectious biological samples.³ These measures affect biobanks on systemic and individual levels from adapting workflows to the use of appropriate personal protective equipment. At BioArkive, our 55+ years of biobanking expertise has enabled us to quickly develop and initiate efficient biosafety procedures for sample handling in the context of the current health crisis.

Challenges related to biobank operations, resources, and infrastructure
The COVID-19 pandemic has impacted transport and delivery of reagents, equipment, and biological specimens needed for the operations of biobanks, and has rendered the operation of biobanks in some geographic areas to maintenance only. Other biobanks have taken on additional projects collecting specimens from patients infected with SARS-CoV-2.  Furthermore, the implementation of epidemiological measures in biobank facilities may have necessitated changes in the organization of their work. Our team at BioArkive offers solutions for global sample distribution, worldwide shipping, and redundant storage locations for business continuity. Moreover, we provide accommodations for regulated and quarantined samples.

Impediments related to the availability of personnel
The COVID-19 pandemic has also affected personnel availability. Some staff members have been unavailable due to an illness or quarantine. There are instances in which biobank employees have had to take on responsibilities related to the collection of samples from patients with COVID-19 and have thus not been able to participate in routine biobank operations. In some cases, epidemiological measures have required employees to work remotely. BioArkive’s team of translational scientists has ensured through thoughtful scheduling and distancing regulations that we are able to continue providing industry-leading cold chain logistics sample storage and professional lab support.

Announcing the launch of our new website Announcing the launch of our new website We are excited to announce the launch of our brand new website at https://www.bioarkive.com/. The official launch will be on September 2nd, 2020. Our goal was to make the new website faster, easier to navigate and more user-friendly.

As San Diego’s leading biorepository and pre-clinical research service provider, it’s important for us to make information regarding our services, capabilities and achievements easily accessible to our current and prospective clients. We endeavor to provide our client partners with the most accurate, up-to-date information and share our knowledge and expertise in the fields of Biobanking, Drug Discovery and Cold-chain Logistics.

It is our desire to provide useful information to our new and existing clients through a non-cluttered, easy to navigate page design. We will post opinions, relevant research findings, and useful reference material through our blog posts. Images and videos posted on our site will give an opportunity for interested parties to come visit us in a virtual setting. Our new website will also act as the access point for a secure client portal, where existing customers can access information about their stored materials. We encourage everyone to visit our social media pages through the buttons provided on the home page.

Pharmaceutical and biotechnology companies spend billions of dollars in drug discovery every year. Despite continuing commitment of extraordinary financial and personnel resources, an alarming proportion of oncology drugs continue to fail during clinical development. A number of causes are likely to account for high clinical failure rates, including (1.) a core discovery focus on preclinical models that poorly reflect complexities of human disease and (2.) an inadequate integration or understanding of translational biomarkers. These shortcomings, coupled with patent expirations, have created substantial revenue pressures. As a result, the pharmaceutical industry has derisked internal research through increased external investments in smaller partner companies who have novel drugs, drug targets or innovative discovery approaches. This approach has already yielded solid results and is exemplified by the emergence of immune targeted agents for cancer, most of which emerged from academic spin-offs and smaller biotechnology companies.

BioArkive has a long (>20 years) track record in working with advanced 3D tumor models that provide novel translational insights on prospective oncology drug candidates. Unlike BioArkive’s tumor microenvironment (TME)-aligned 3D models, the industry standard for drug screening, more often than not, inadequately mirrors relevant human tumor physiology. Some examples of poorly modeled attributes of human cancer include:

• Oxygen: ≤ 4.5% (blood capillary with tumor gradient)
• pH: pH=6.5-6.9 (acidified TME; Warburg Effect)
• Stiffness: ≤ 5,000 Pa (vs. 10^9 Pa for 2D)
• Dimensionality: Three dimensional (3D) environments
• TME Complexity: Complex, dynamic, limited nutrients
• Glucose: Limiting (less than 8-9 mM)
• Immune System: Inflammatory (cytokines, growth factors)
• Early Disease: 1st or 2nd line vs. late – multiple relapsed/refractory
• Tumor Cell Division: Weeks to months (human condition vs. day(s) in 2D)
• Cell-cell interactions: Extensive, multicellular interfaces