Lecchain

Thermotolerant and oral vaccine candidate for COVID-19 based on plant lectins that does not require cold chains for maintenance

Our solution is a subunit, oral and thermostable vaccine against COVID-19 called Lecchain. The antigen is expressed in an Escherichia Coli strain and composed of the receptor-binding domain (RBD) peptide of the S1 protein from the SARS-CoV-2 virus fused to the plant lectin Ricin Toxin B (RTB). This chimeric protein (RTB::RBD) was designed to improve thermal stability of the RBD antigen due to the properties of plant lectins. Additionally, it allows the exchange of this part by other antigens, making Lecchain a modular system that could target other diseases. The recombinant chimeric protein will go through thermal-stability assays and a final immunological assay based on an Elisa for evaluating the proper tertiary structure integrity of the RBD. Unlike current commercial vaccines, Lecchain is a vaccine candidate intended to be delivered in absence of cold chains, tolerating high temperatures and manufactured in recombinant expression systems scalable in inexpensive bioreactors, generating minimal waste.

Distribution and storage of vaccines in low-income countries are difficult and expensive. The current outbreak of the novel coronavirus SARS-CoV-2 has paid a high toll in terms of human cost, reaching now almost 326 million confirmed cases and more than 5.54 million global deaths. Complex logistics, distribution, and storage are needed for these vaccines but the poor health systems of some countries are huge barriers to achieve complete immunization. COVID-19 vaccines that have demonstrated better efficacy results need lower temperatures for their preservation. Moreover, it is reported that 80% of the vaccination costs are due to the cold chain. 

Likewise, according to The Economist Intelligence Unit, around 108 countries wont have widespread vaccination access until 2023. Rural communities have severe risks due to lack of vaccine cold chains. Local statistics show that while the capital state (Lima) has around 78% of vaccinated people, poorer and more rural states like Puno, Madre de Dios and Loreto have lower than 50% vaccination rates. This gap affects millions of people not only in Peru but in other underdeveloped countries in the world.

We classify our initial target population in three main aspects: demographic, location, and psychographic. 

Demographic: 18+ years old, comorbidity, Peruvian, low incomes

Location: Hard to reach places, outside Lima, Region of Lima, Varied weather. 

Psychographic: The nearest health center is 4-10 hours away, they hardly ever see a nurse or a doctor, they do not get vaccinated with frequency and they have not received a COVID-19 dose or just once. 

Thanks to Lecchain, people from last-mile places will have access to vaccines without the need for cold chains for their maintenance. This will improve the speed of the vaccination process in the country for the following diseases that require doses and the users will no longer be concerned about the efficiency of vaccines. Likewise, health professionals will no longer carry out all the vaccines in cold boxes for a limited time before they get lost, walking for long hours or traveling by boat to reach every community.

We have developed a framework of 3 phases: identify, invent and implement 

• As a team we have engaged with potential users for the first stages on our project. Main insights and needs were identified from interviews along with medical professionals and local health authorities that work in rural areas of Peru. We also identified the different concerns from the public, while they were getting vaccinated in a local vaccine distribution center and afterwards in our social media platforms. 

• Vaccination experts such as MASED, Roche and Genlab specialists helped us ideate by sharing with us that a good solution must have thermostability, be cheaper than the current logistic system and vaccines must have a high immunogenicity. Helped by these interactions we optimized the solution multiple times during the last year, as such, our product was adapted continuously according to what we found when we approached the population. 

• That said, the objective is to design and implement an innovative vaccine formula for COVID-19 that can withstand high temperatures without losing its efficacy through recombinant protein technology and plant lectins, which would facilitate its adaptability for other diseases.

The development cycle of our solution consists of four main stages: DESIGN, BUILD, TEST, and LEARN. Until now, we have focused on the DESIGN phase. First we identified the problem, it implied being aware regarding the concerns and problems around vaccination and cold chains. It was followed by doing deep research around topics such as vaccines types, immunological aspects of the disease, thermotolerant vaccines, and recombinant technologies to name a few.  As well as, talking with different health professionals, scientists, biologists to gather insights to validate our solution.

The next steps were focused on the optimization in silico of the composition of the vaccine, which meant to work on the computational part of the solution. For that, we used different protein modeling software to evaluate several conformations of the vaccine at different conditions. This helped us to evaluate the immunogenicity, antigenicity, and thermal properties of the proposal vaccine. All this work took us to participate and be evaluated in the iGEM Design League competition and won the Gold Medal, Best Human Practices, and SDG Impact awards. Lecchain has been proposed and optimized thanks to feedback from professionals in the area of health, biotechnology, and by the user itself. 

Lecchain is a bacterial cell-expressed subunit, oral and thermotolerant vaccine against COVID-19. Powered by plant lectin-based technology, this vaccine can be transported without a cold chain system, which ensures a fast and efficient delivery of this product in rural and remote communities. The double function as an adjuvant and thermoprotector of RTB lectin confers the immunogenicity and protective efficacy required. 

Excluding COVID-19, millions of people are still dying because of poor vaccination systems in their countries. The programmable design of Lecchainthrough synthetic biology enables it to combat several diseases thanks to its modularity to assemble different types of antigens. Therefore, Lecchain will help in last-mile applications for temperature-sensitive vaccines and deal with emerging pandemics

Unlike many current vaccines which are expressed in mammalian cells, Lecchain is produced in E. Coli. It is easy and inexpensive to produce our protein in this bacteria at large scales because fermentation is already an established process in the biopharmaceutical industry. Besides, the production of recombinant vaccines has less risks when compared to other forms since it does not require cultivation of dangerous infectious agents.

According to the latest study by The Economist Intelligence Unit (EIU), it was estimated that about 108 countries would not have widespread vaccine access until 2023 or more years. Within those countries, people living in rural areas can be expected to face more severe risks due to lack of access to vaccine cold chains. Thus, an estimated 1.42 billion people living in the last mile in these countries are at high risk, even if only temporarily, due to lack of access to refrigeration that prevents them from accessing the COVID-19 vaccine.

After completion of the test phase, our solution plans to initially benefit the Peruvian populations of San Martin, Huanuco, Huancavelica, Amazonas, Ayacucho, Ucayali, Madre de Dios, Loreto and Puno, which are awaiting vaccines. These regions have the worst vaccination rates (max: 59.9%, min: 45.8%) together with poor development in infrastructure and transportation. So, once our solution is launched it can help potentially 2,924,588 people in these regions who are still not receiving any doses to have access to vaccination.

For this year we are prepared to start the experimental part of our solution. In that sense, our impact goals are focused on this aspect. We have determined two of them in the short term:

• Prove thermotolerance characteristics of the chimeric antigen RBD-RTB under different temperature tests.

How? Thermotolerance assays and SDS Page evaluations of the protein will be executed

• Evaluate immunogenicity of the chimeric antigen 

How? In vitro immunological assays such as ELISA will be performed to evaluate tridimensional conformation of the protein after being exposed to different temperatures.

In terms of the long term goal, our main objective is to 

• Immunize people that live in far away communities and who have not received any vaccine yet.

How? Vaccines will be distributed mainly to far away communities throughout a series of agreements with the government

1. Prove thermotolerance characteristics of the chimeric antigen RBD-RTB under different temperature tests.

• Range of temperature without denaturation

• Percentage (%) of temperature difference that LECCHAIN supports in contrast to current vaccines

2.  Evaluate immunogenicity of the chimeric antigen

• Efficacy and effectiveness rates

 3. Immunize people that live in far-away communities and who have not      received any vaccine yet

• Proportion of population vaccinated for the first time in rural and remote communities

• Number of people requiring interventions against COVID-19 in these areas

The main barriers belong to the financial, legal, and cultural areas. About the financial side, we need funding support to cover expenses to develop the experimental phase. Although we currently have the support of some institutions that guarantee our project, having a fund that gives us the economic capacity would help us develop our project with the best results.

In the legal area, in Peru, there are no specific limitations that avoid the development of the proposal based on synthetic biology at the laboratory level, so it does not represent a barrier during the entire experimentation process. However, there is still little development and promotion of biotechnological proposals in the health sciences. Therefore, if it is decided to take the proposal to experimentation on a larger scale, it would have to go through a series of bureaucratic processes that would slow down the approval process of experimentation phases.

In the cultural area, the limitation would come from the fear that the population may have regarding the use of new biological technologies such as synthetic biology or anti-vaccine movements. This limitation originates mainly from the lack of knowledge of these new areas.

Being a synthetic biology solution, the team is made up of 12 bioengineering students with several backgrounds. We are university students with knowledge in the area and with experience in research work, wet lab, and dry lab. Lecchain seeks to enhance its talents to maximize the results of our solution.

Marcelo, the leader, is in charge of planning and guiding the project. He seeks to develop inclusive work and highlights the team's achievements. Gladys and Jose have experience in wet lab topics such as recombinant protein expression. Marcelo and Nathaly have experience working in bioinformatics projects and they are in charge of developing the computational modeling of the protein. We are also advised by a professor who is a biotechnologist with experience in recombinant systems using plants, who counsels us on different topics to improve the solution.

Committed to delivering the solution to all the people, we have a marketing team (Fatima, Massiel, and Allison) who is in charge of communicating the project to the general public. And a technology area by Ariana and Alessandro in charge of developing Lecchain's website. 

Our members from the human practices area Luis, Diana, Amalia, Ximena y Yomali have been in charge of gathering information by meeting, talking, and understanding the needs with specialist doctors, nurses, health center workers, researchers, users in order to validate our problem and receive feedback on our solution. These dialogues have allowed us to form ties with companies in the health sector and obtain funding for materials needed to carry out the project. 

We are last-year students from Bioengineering. Each of us has experience working in areas like synthetic biology, tissue engineering, bioprocesses, bioinformatics, and microfluidics to name a few and we are committed to gathering all this knowledge for developing a good solution.

We have partnered with two biotechnology companies: Roche, Mased, GenLab and Biotech. They gave us valuable technical and management feedback on the solution we proposed through virtual meetings. They also financed the purchase of the reagents necessary for the experimental development.
We formed alliances with internationally certified student organizations focused on the biotechnology field: SBE-UTEC and Piso 11.

• With the SBE-UTEC and its Genetic Engineering subcommittee, we participated in dissemination activities with participants from several Latin American countries. From this meeting we were able to understand the perspective regarding synthetic biology of the people in the region.

• Our alliance with Piso 11 was evidenced through several activities on social networks. An interview was conducted with members of the team, in order to make the project better known within our university community. It also allowed us to know the doubts of people who are not part of the project.

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Lecchain is a good candidate for the Pozen Social Innovation Prize, because it arises as a proposal to mainly contribute to the achievement of sustainable development goals No. 3, 9, and 10. 

Lecchain gets along with the achievement of objective N° 3 of health and well-being since we seek to guarantee the healthiness of all people through the development of a candidate vaccine against COVID 19. This is because we are currently facing a global health crisis of COVID 19 that has caused millions of deaths and has brought about destabilization of the global economy. In addition, it has intensified existing inequalities and has well affected the poor and the most remote and vulnerable communities. Therefore, the development of a vaccine could help reduce the gaps in access to health systems in many poor countries such as Peru.

Our proposal is also aligned with SDG 10 to reduce inequalities since we seek to close gaps and reach the most remote cities of the country without the need for a complex and expensive cold chain. Taking into account that the role of rural women has not yet reached the same conditions and opportunities as rural men or women in urban areas. Lecchain is an opportunity to break this cycle of inequalities, so that girls and women can exercise their rights to a dignified life through access to vaccines.

In the design, optimization, and testing of our proposal, we align ourselves with objective N° 9 of industry, innovation and infrastructure. Lecchain seeks that the design is optimized for easy and feasible mass production because we propose to introduce and promote new technologies such as synthetic biology and allow efficient use of resources, where women can disrupt this new market. 

With the additional funding from the prize, we would apply Lecchain technology to design new vaccines against SARS-CoV-2 variants and other common diseases. In addition, we could start the development of the preclinical trial phase of the Lecchain vaccine candidate in order to get information about its efficacy, toxicity, and pharmacokinetic properties before human trials.