Nanobodies Production.docx

“Nanobodies production” By Cintia Torres (6324452) According to the World Health Organization (WHO) (1), with more than 3.7 million new cases and 1.9 million deaths each year, cancer is the second most important cause of death in Europe. Globally, cancer accounted for 8.2 million deaths (around 13% of the total) in 2012. The great impact of this disease in developed and developing countries makes it a very interesting topic to research on how to appropriately treat it. One of the main problems of the anticarcinogenic therapy used nowadays is the very strong side effects developed because of the lack of specificity of many drugs to fight cancerous cells. Nanobodies used as a messaging system have a great potential in solving this problem. The explanation of what nanobodies are and what is their potential use in target therapy for cancer were discussed in the essay “Nanobodies for targeted therapy” based on the lecture of Dr. Paul van Bergen en Henegouwen. In this second essay, the production of the nanobodies will be explained and thus, the research question: what steps should be taken to produce stable and specific nanobodies? will be addressed. As explained in the before mentioned essay, nanobodies are basically obtained from animals such as llamas and alpacas. The process to obtain a complete drug delivery system starts from this point. In summary, the steps taken to obtain such specific messaging system comprises: 1. Identifying a target tumour 2. Immunization of llamas and recovery of the produced nanobodies 3. Nanobody selection 4. Nanobody characterization 5. Functionalization 6. Imaging 7. Therapy (in animal model). After the tumour of interest has been identified, a proper biomarker for this tumour should be selected. Then, the actual production of the nanobody starts by immunizing the llamas. After two months, half a litre of the blood is collected from the animals and the DNA from their cells is purified. From the RNA, an enrichment of the sequence that makes the heavy chains is carried out. Then by a “phage selection” process, the specific nanobodies are isolated and amplified using host bacteria. After the binding of the nanobody is confirmed and characterized, a step of functionalization to bind the therapeutic molecule is carried out using “click chemistry”. Imaging by an optical analytical technique is performed to confirm the binding of the antibody and the drug molecule, followed by the addition of a linker. The last step is the therapy using animal models. References (1) World Health Organization Europe. Data and statistics. (2018). Retrieved from http://www.euro.who.int/en/health-topics/noncommunicable-diseases/cancer/data-andstatistics