Safety and Security

Project Overview

The objective is to design a biosensor for the early detection of Amyotrophic Lateral Sclerosis (ALS), a neurodegenerative disease that nowadays doesn't have a diagnostic method.

The proposed system is based on the use of aptamers, short DNA sequences capable of recognising and binding specifically to biomarkers. In our case, the objective is to detect 2 isoforms of 2 proteins; "Protein A" and "Protein B" generated due to the aberrant splicing of their genes. The splicing change is due to the dysfunction of the splicing factor TDP-43, the main cellular hallmark of ALS (Seddighi et al. 2024). The aberrantly included exons are called cryptic exons, which give rise to cryptic peptides (CPs) in the respective proteins. A CP is therefore an extra amino acid sequence found within (between) the normal protein. Both proteins are primarily found in the cerebrospinal fluid. The aptamer will be designed to recognize exclusively the extended form of the protein containing the CP, and not its normal form.

For the development of the project, cloning will be performed using the TrcHis B plasmid. The four fragments corresponding to the human variants of the proteins of interest, namely the normal and extended forms, will be inserted into this plasmid. The proteins will be expressed and purified in Escherichia coli, obtaining both the normal and extended forms (which include the PC) of Protein A and Protein B.

These proteins will subsequently serve as models for the design and evaluation of specific aptamers. Experiments to obtain and characterize the aptamers will be carried out to validate their specificity and applicability in the biosensor system.

Research Team Data

Potential Risks

Equipment Required

Chemicals Used

Chemical burns or irritation may result from strong bases like sodium hydroxide or acids like boric acid, as well as from detergents like SDS and Triton X-100.

Toxicity or systemic effects could arise from reagents such as ethanol, methanol, β-mercaptoethanol, DTT, and ethidium bromide if inhaled, ingested, or absorbed through the skin.

Allergic reactions, respiratory issues, or long-term health effects may occur from repeated exposure to enzyme inhibitors (PMSF, leupeptin, pepstatin A, benzamidine) or antibiotics (ampicillin, chloramphenicol), while acrylamide carries a neurotoxic and potential carcinogenic risk.

Biological Agents

Safety Measures

To ensure safe work in the lab and minimize risks, we follow these documents:

• Biological agents and waste: Document waste
• General lab rules: Document general rules
• Legislation: Document legislation
• Risk prevention:Document risk prevention

Basic Lab Information

Data Protection & Confidentiality

In the development of the ALSense project, we have adopted specific measures to ensure that all information collected in interviews, workshops, and meetings with experts, associations, and potential patients is managed in accordance with the highest standards of data protection. This commitment is grounded in the applicable legal frameworks: the General Data Protection Regulation (Regulation (EU) 2016/679, GDPR) and the Spanish Organic Law 3/2018 on the Protection of Personal Data and Guarantee of Digital Rights (LOPDGDD).

Principle of data minimization

According to Article 5(1)(c) of the GDPR, personal data must be "adequate, relevant and limited to what is necessary in relation to the purposes for which they are processed." In our project, this meant collecting only information relevant to the research objectives: perceptions about the biosensor, ethical concerns, and suggestions for improvement. At no point were complete medical records or unnecessary sensitive data requested.

Transparency and informed consent

Article 5(1)(a) of the GDPR establishes that data must be processed "lawfully, fairly and in a transparent manner," while Article 6 specifies that one of the legal bases is the explicit consent of the data subject. In practice, before each encounter, participants were provided with a clear informed consent form explaining the purpose of data collection, how the information would be used, and their right to withdraw at any time. In meetings involving sensitive content (e.g., patent-related issues or technical aspects of the biosensor), participation was restricted to authorized members, reinforcing the security of shared information.

Protection against risk and restricted access

Article 32 of the GDPR requires that appropriate technical and organizational measures be implemented to ensure a level of security appropriate to the risk. In ALSense, only specific team members had full access to meeting records and interviews. Whenever the information required greater confidentiality, strict access restrictions were applied.

Protection of health data

Article 9 of the GDPR and Article 9 of the LOPDGDD state that health data is a specially protected category and may only be processed with a valid legal basis and adequate safeguards. Therefore, no direct clinical data or identifying information from patients was ever collected. In surveys directed to patients and families, participants were always given the option to respond anonymously, creating a safe and pressure-free space to share their views.

Rights of participants

Article 13 of the LOPDGDD guarantees participants the rights of access, rectification, erasure, and objection. In line with this, all participants were informed of these rights and of the possibility of exercising them at any time.

Proactive responsibility

Article 24 of the GDPR introduces the principle of accountability, requiring data controllers to demonstrate compliance with the regulation. In the ALSense project, every meeting, consent form, and decision on data handling has been documented, ensuring full traceability and transparency throughout the process.

Every step in handling information has been carefully designed to meet the highest safety standards, ensuring both the protection of participants and the integrity of the research environment.

For more details on the consent process and the rights granted to participants, you can access the Consent Form Document here:
Documento General de Consentimiento Informado (ALSense-UIC).pdf

References

Aljohani, Maher M., Dana Cialla-May, Jürgen Popp, Raja Chinnappan, Khaled Al-Kattan, and Mohammed Zourob. 2022. “Aptamers: Potential Diagnostic and Therapeutic Agents for Blood Diseases.” Molecules (Basel, Switzerland) 27 (2).

Harper, Sandra, and David W. Speicher. 2011. “Purification of Proteins Fused to Glutathione S-Transferase.” Methods in Molecular Biology (Clifton, N.J.) 681:259–80.

Seddighi, Sahba, Yue A. Qi, Anna-Leigh Brown, Oscar G. Wilkins, Colleen Bereda, Cedric Belair, Yong-Jie Zhang, et al. 2024. “Mis-Spliced Transcripts Generate de Novo Proteins in TDP-43-Related ALS/FTD.” Science Translational Medicine 16 (734): eadg7162.

Smith, D. B., and K. S. Johnson. 1988. “Single-Step Purification of Polypeptides Expressed in Escherichia Coli as Fusions with Glutathione S-Transferase.” Gene 67 (1): 31–40.

Zhao, Jiayao, Jian He, Liping Zhong, Yong Huang, and Yongxiang Zhao. 2020. “Production of Aptamers by Cell-SELEX and Their Applications in Cancer Biomarker Identification.” Discovery Medicine 29 (158): 159–67.

Unión Europea. (2016). Reglamento (UE) 2016/679 del Parlamento Europeo y del Consejo, de 27 de abril de 2016 (Reglamento general de protección de datos). Diario Oficial de la Unión Europea. https://www.boe.es/doue/2016/119/L00001-00088.pdf

España. (2018). Ley Orgánica 3/2018, de 5 de diciembre, de Protección de Datos Personales y garantía de los derechos digitales. Boletín Oficial del Estado, núm. 294. https://www.boe.es/buscar/act.php?id=BOE-A-2018-16673