Denomination: Therapeutic potential of recombinant antibodies to neutralize Immunogenic peptides in celiac disease — GIPTRAP.
Start Date: 01/2022
End Date: 01/2023
Trainee Researcher: JIMÉNEZ FERNÁNDEZ, ALICIA
Celiac disease is triggered by gluten consumption and causes atrophy of the intestinal villi, resulting in poor absorption of nutrients. The main sources of gluten are wheat, rye, barley and some types of oats. At present, the only effective treatment is a gluten-free diet and, consequently, excluding the consumption of wheat, rye, barley and oats, something difficult to achieve. Preliminary data revealed that an antibody against the most immunogenic gluten peptide suppressed 90% of the ‘ex vivo’ immunogenicity of hydrolyzed gluten. The EU-funded GIPTRAP project aims to develop a therapeutic strategy to overcome the effect of gluten exposure in celiac patients. The objective is to assess the potential of antibodies againts gluten immunogenic peptides as therapeutic tools, evaluating their “in vitro” and “ex vivo” effectiveness.
Celiac disease consists of an autoimmune response triggered by dietary gluten exposition in genetically predisposed individuals. To this day, the only effective treatment is a permanent gluten free diet (GFD). However, a fully GFD is difficult to achieve, and symptoms persist in 65% of patients, with only 8% of them reaching duodenal-mucosa complete normalization. In collaboration with international research institutions, Biomedal S.L. showed that at least two thirds of celiac patients consume gluten aware or unaware at least once per week, and only 11% do not ingest gluten in a period of four weeks.
GIPTRAP aimed at evaluating the potential use of antibodies against gluten immunogenic peptides (GIP) as therapeutic tools to reduce the effect of exposure to gluten in celiac disease patients.
GIP neutralization capabilities of several antibodies were tested in vitro in an ELISA sandwich. Biomedal S.L. licensed G12 antibody showed the best results, reducing 97% of GIP signal. Neutralization still needs to be tested ex vivo to confirm that this GIP-capture does not induce celiac disease patient T-cells activity. An ethical committee has been obtained approving the protocol to purify T-cells from blood samples from healthy volunteers and celiac disease patients. Currently, samples are being collected and ex vivo assays will be carried out before the end of the year by Biomedal S.L. and collaborators.
Antibody G12 engineering has been carried out. Recombinant, Fab and scFv versions of G12 were generated and assessed for anti-GIP activity. The recombinant version was as efficient as the original. However, when Fab and scFv versions were used in an ELISA test, they were not as sensitive as G12. The scFv version will be used for antibody maturation by phage display before the end of the year by Biomedal S.L. and collaborators.
Antibodies have been produced using hybridoma cells, CHO and HEK mammalian cells, and E. coli bacterial cells, all in batch cultures. A hollow-fiber continuous culture system was acquired to enhance production yield. It will be used for matured antibody production, but still needs to be optimized.
In relation to project dissemination, Alicia participated in two life science student events at Universidad Pablo de Olavide to discuss career opportunities and promoted MSCA-IF-SE fellowships. She attended PEGS Europe 2022, a summit in Protein and Antibody Engineering, and Trasnfiere2023, a tech transfer trade fair organized in Malaga, Spain.
Alicia has also completed a training program that has equipped her with tools to follow and draft legal documents related to intellectual property, use R-Studio for multiple data analysis approaches, comprehend in vitro diagnosis and quality management regulation (ISO13485) and work within its framework, and improve her skills in project management and science communication. All this achieved in certified courses.
To sum up, accumulated results indicate that ex vivo antibody GIP-neutralization is feasible, especially if a matured antibody specifically design with this purpose is obtained. However, transferring this proof of concept into therapy will be a challenge in terms of production, delivery, clinical ethical clearance, and regulation. Still, tools and know-how produced during this project will help advancing celiac disease diagnostic and research.