Periodic Reporting for period 1 - PRe-ART-2T (Predictive REagent-Antibody Replacement Technology stage 2-Translation)
Período documentado: 2022-06-01 hasta 2023-05-31
Antibodies are one of our major lines of defence against infection and our bodies can create them very quickly to help incapacitate a multitude of biological invaders. Humans do this by changing the part of the antibody that recognises the invading pathogen, though a process of rapid, natural mutation. Antibodies are also used in medicine and in scientific research and both PRe-ART and it’s follow on project PRe-ART-2T focus on the latter – antibodies for use in scientific research.
It is really important to understand the difference between antibodies used as medicines, known as “therapeutic antibodies” and those that are used in research. Like any other medicine, therapeutic antibodies are very tightly regulated and are exceptionally expensive. Their extremely high price results from the massive research effort involved in creating them, characterising them (understanding their structure, what they do and how they work) and in quality control - making sure that each batch of these very complicated molecules is the same. That all comes before the enormous cost of clinical trials that are required to determine safety and efficacy (whether or not the antibody actually works to treat the disease or condition in question) before such antibodies can ever go into humans as medicines.
In contrast, research antibodies, otherwise known as “reagent antibodies”, are never injected into people. Rather, they are used to detect proteins of interest in fundamental biological research and in diagnostics. Originally, research antibodies were generated mimicking the natural process – by injecting an animal with the protein target of interest, taking a blood sample and purifying the resulting antibodies. However, that is relatively rare now. For the last forty years, biological research has relied on monoclonal antibodies (mAbs) to detect biomolecules within medical diagnostics and basic life science research. Without them, detection of disease-state biomarkers would be impossible. Yet unfortunately, it has become very evident that many mAbs are not nearly as specific as presumed, many are poorly characterized and their production is based on unstable cell lines. In fact, about half of the commercial reagent mAbs have been shown to not function correctly. This equates to a wastage of ~$1 billion of research money worldwide each year - a very significant proportion of which occurs in the EU. Moreover, that cost is purely of the antibodies themselves. The waste of money and time on experiments that fail, or worse, experiments that appear to have worked – only to find that they are wrong or can’t be reproduced, is unfathomable. The status quo continues because research antibodies are relatively cheap (typically around €300 per sample rather than tens of thousands of Euros for therapeutic antibodies). Research is very rarely funded to the level where purchase of therapeutic-grade antibodies is remotely possible, and so the money just isn’t there to justify the suppliers to make lots of high-quality research antibodies.
The purpose of PRe-ART and Pre-ART-2T is to solve this problem. We are creating a radically new process to make synthetic antibody replacements that have the quality of therapeutic antibodies at the price of standard research antibodies, so that all researchers – academic and industrial, can afford high quality research antibodies and thus eliminate such terrible wastage.
PRe-ART developed the contributing technologies, while PRe-ART-2T is completing that technology, testing it in our own and industrial laboratories and then starting a company to make these antibody replacements available to all research scientists. We estimate that if we captured even 5% of the research antibody market, we could save the EU in the order of €70 million per annum.
It is really important to understand the difference between antibodies used as medicines, known as “therapeutic antibodies” and those that are used in research. Like any other medicine, therapeutic antibodies are very tightly regulated and are exceptionally expensive. Their extremely high price results from the massive research effort involved in creating them, characterising them (understanding their structure, what they do and how they work) and in quality control - making sure that each batch of these very complicated molecules is the same. That all comes before the enormous cost of clinical trials that are required to determine safety and efficacy (whether or not the antibody actually works to treat the disease or condition in question) before such antibodies can ever go into humans as medicines.
In contrast, research antibodies, otherwise known as “reagent antibodies”, are never injected into people. Rather, they are used to detect proteins of interest in fundamental biological research and in diagnostics. Originally, research antibodies were generated mimicking the natural process – by injecting an animal with the protein target of interest, taking a blood sample and purifying the resulting antibodies. However, that is relatively rare now. For the last forty years, biological research has relied on monoclonal antibodies (mAbs) to detect biomolecules within medical diagnostics and basic life science research. Without them, detection of disease-state biomarkers would be impossible. Yet unfortunately, it has become very evident that many mAbs are not nearly as specific as presumed, many are poorly characterized and their production is based on unstable cell lines. In fact, about half of the commercial reagent mAbs have been shown to not function correctly. This equates to a wastage of ~$1 billion of research money worldwide each year - a very significant proportion of which occurs in the EU. Moreover, that cost is purely of the antibodies themselves. The waste of money and time on experiments that fail, or worse, experiments that appear to have worked – only to find that they are wrong or can’t be reproduced, is unfathomable. The status quo continues because research antibodies are relatively cheap (typically around €300 per sample rather than tens of thousands of Euros for therapeutic antibodies). Research is very rarely funded to the level where purchase of therapeutic-grade antibodies is remotely possible, and so the money just isn’t there to justify the suppliers to make lots of high-quality research antibodies.
The purpose of PRe-ART and Pre-ART-2T is to solve this problem. We are creating a radically new process to make synthetic antibody replacements that have the quality of therapeutic antibodies at the price of standard research antibodies, so that all researchers – academic and industrial, can afford high quality research antibodies and thus eliminate such terrible wastage.
PRe-ART developed the contributing technologies, while PRe-ART-2T is completing that technology, testing it in our own and industrial laboratories and then starting a company to make these antibody replacements available to all research scientists. We estimate that if we captured even 5% of the research antibody market, we could save the EU in the order of €70 million per annum.
Research antibodies are obtained in one of three ways. If an antibody already exists, it can be bought from a catalogue. Some catalogue antibodies have been tested for specificity (depending on the supplier), but many have not. Alternatively, some antibodies are “recombinant”. These are discovered by screening a “library” – a mixture of typically billions or trillions of synthetic antibodies made in the laboratory without any animal involvement. These are usually better characterised, but there are comparatively few available. If there is no catalogue antibody available, a “custom antibody” has to be made by one of these three routes. This requires the target protein to be available, purified and then used either to raise a polyclonal or monoclonal antibody or to screen a synthetic library. Generating a custom antibody is both time consuming and expensive.
We are developing an entirely different process. In PRe-ART, we merged life sciences, protein engineering and computational prediction into a combined technology to create antibody alternatives in lots of individual modules – each binding one component of a protein called an amino acid. Conceptually, these modules can be likened to Lego bricks. In PRe-ART-2T, we are completing our set of Lego bricks (imagine a whole palette of different coloured bricks) and learning how to combine them together properly. To continue the analogy, imagine, as a child, building a Lego brick wall. If you simply build bricks on top of each other, the wall will be unstable, but if you overlap them together properly, the wall will be strong. In the first year of PRe-ART-2T, we are learning how to build our bricks together properly, so that the scientists can have any combination of “colours” in whatever order they need. So far, the results are very good – we are learning a lot about combining bricks effectively, and the walls that we have built to date are strong. In scientific terms we are learning how to combine the modules and we have bound initial target sequences with both high affinity and specificity. The work will continue in years 2 and 3 of the project. At the same time, we are scoping a business, so that we can supply our antibody replacements worldwide to supply researchers with cheap, effective and stable antibody replacements.
We are developing an entirely different process. In PRe-ART, we merged life sciences, protein engineering and computational prediction into a combined technology to create antibody alternatives in lots of individual modules – each binding one component of a protein called an amino acid. Conceptually, these modules can be likened to Lego bricks. In PRe-ART-2T, we are completing our set of Lego bricks (imagine a whole palette of different coloured bricks) and learning how to combine them together properly. To continue the analogy, imagine, as a child, building a Lego brick wall. If you simply build bricks on top of each other, the wall will be unstable, but if you overlap them together properly, the wall will be strong. In the first year of PRe-ART-2T, we are learning how to build our bricks together properly, so that the scientists can have any combination of “colours” in whatever order they need. So far, the results are very good – we are learning a lot about combining bricks effectively, and the walls that we have built to date are strong. In scientific terms we are learning how to combine the modules and we have bound initial target sequences with both high affinity and specificity. The work will continue in years 2 and 3 of the project. At the same time, we are scoping a business, so that we can supply our antibody replacements worldwide to supply researchers with cheap, effective and stable antibody replacements.
Everything to date is “beyond the state of the art” since no one else is creating a modular alternative to antibodies. If successful, we will revolutionise the whole process of obtaining a research antibody. There will be no need to purify the target protein – only the amino acid sequence, via email, will be required. There will be no animals involved, no mAbs and no screening of synthetic libraries. We will simply assemble the encoding DNA from pre-defined modules, express the gene and ship the resulting “antibody” to the researcher. Since the process will ultimately be automated, production will be both very rapid and cheap. Most importantly though, our antibody alternatives will do exactly what they are meant to do.
It should be noted - our antibody replacements won’t work for everything. They bind linear peptide sequences rather than three dimensional shapes - though, in research, many such linear sequences exist.
It should be noted - our antibody replacements won’t work for everything. They bind linear peptide sequences rather than three dimensional shapes - though, in research, many such linear sequences exist.