The Reid Bioanalytical Forum was established in 1973, initially starting life off as the Bioanalytical Forum, and with the untimely passing of Professor Eric Reid, a long-time organiser of the event, the name was altered. It is held every other year and this year was the 23rd running of the event. The focus of the event is very much bioanalysis, and so is not devoted just to mass spectrometry, chromatography, ligand binding assays or regulatory aspects, but instead covers the full breadth of technical and regulatory aspects that relate to the analysis of a biological sample. The ethos of the meeting is very much about full engagement of the audience, which is achieved through honest presentations and also an active social agenda during the three days over which it is held. This year’s meeting held true to this ideology, with a full list of presentations and also social activities.
The 23rd Reid started off in usual fashion with an interactive social event. On this occasion a pub quiz, chaired by Tim Sangster. Most of the delegates had already started to fuel up and so with everybody in good spirits much merriment was to be had under the direction of the Symposium chair. which was an amazing success. There were many highlights of the evening including the amazing charades round which unveiled a very different side of Sangster and Staelens, one which was probably best not revealed in such a public manner. The winners of the quiz demonstrated their lack of a well spent youth, with an unfeasibly large percentage of correct answers to a very broad set of questions.
Winners of the evening quiz
The first day proper of the event start at 8-30, perhaps a little early for some of the delegates given the nature of the quiz celebrations the previous evening. The chair for the first session was Tim Sangster. He welcomed the delegates to the 23rd Reid, and discussed the ethos of the Reid, which is about engagement, and he tasked the delegates to ensure that they get to know all of their counterparts. He then proceeded to introduce the first speaker. There was a change due to Professor Barran from Manchester not being available, however Professor Wilson from Imperial College stepped into the brink to give one of his classic lectures on metabolomics.
Ian Wilson (Imperial College)
Criteria for accepting the data obtained from multi-analyte biomarker and drug bioanalysis
The first speaker was introduced as the last man standing of the original Reid organising committee. Professor Wilson Ian started his presentation by reiterating the ethos of the Reid, before moving onto his research areas of phenotyping. He described the approach was initially to start off with untargeted analysis, stressing that it was not just about finding a specific biomarker but more of a flavour of what was happening in the biological system. Ian then moved to describe targeted analysis.
Ian stated that his work was focussed in the discovery part of the drug development program, doing good science but not in a regulated environment. He introduced the biochemicals pathways associated with the metabolome, stating that this was far too complicated to fully understand but then stated that his research was more looking at a small area of this. With a focus on targeted assays, the speaker started to discuss some of the validation criteria. He stated that he was not sure what was actually required. He gave an example of tryptophan and its catabolites, stating that at Imperial he and his team had developed an assay for this which met the FDA validation requirements.
He showed the chromatography, demonstrating one of the issues which was the very broad range of polarities and hence a range of retention times. He also stated that it was not possible to have a blank matrix and so the assay was developed was using water as the matrix, which clearly is not representative of what the matrix is. He presented the figures of merit which were very impressive, with the specifications stipulated by the FDA being met for all of the compounds.
One of the issues that Ian demonstrated was that on occasion he was monitoring very different ranges of compound concentrations, and this resulted in trying to detune the mass spectrometer to ensure that all of the components can be seen on the same scale. The use of a pooled standard ensures that the assay has greater stability.
Professor Wilson then moved on to discuss the possible outcomes associated with the analysis of 18 analytes. He looked at the extremes where analytes all pass or fail, and then moved into what happens if one of the analytes does not pass but 17 do, and also looking at the converse where 17 analytes fail but one passed. He asked the audience the question at what point would the assay fail, in terms of the number of passed analytes, and also the type of failure that was incurred. Ian wrapped up this part of the session by stating that the rules governing multi analyte validation were still being developed.
Ian then moved the discussion onto exogenous drugs, where matrix can be used. He gave an example of paracetamol, with its metabolites again asking the same questions regarding what to accept as the validation criteria. He stated that one of the issues when developing an assay is that typically the dynamic range is not known before the samples are taken. This sparked some debate regarding the nature of the validity of extrapolating the data.
Ian then wrapped up his lively presentation stating that for multitarget analysis there is rarely a digital response for if the assay can be validated. He stated that there has to be a degree of pragmatism associated when looking at sample data that does not meet the validation criteria.
Sue Walker & Rebecca Mogey
Utilising SPE for the collection and storage of faecal hormone metabolites – Invaluable physiological data for endangered species in the field
Sue started of introducing herself as a reproductive biologist stating that she was not a bioanalyst. She then moved onto describing what a zoo does, and in particular looking to try to address the loss of biodiversity that is happening in the world. She presented data which showed the very dramatic impact that humans have had on the extinction of animals, stating that this had increased by a factor of 50 due to human influence. With 700 million visitors, zoos make an ideal vehicle for educating the public, as well as the potential to generate $1 billion in revenue to support conservation. This has been driving investment, coupled with the depth of scientific knowledge and the connections with the relevant NGO and governmental organsations.
Sue then moved onto what the mission statement was for the zoo stressing the conservation aspect and their charitable status. With a team of 14 scientists they look to provide evidence for the impact that humans have on the animals, so that informed decisions can be made. Sue proceeded to give an example of the eastern black rhino research. With a world-wide population of 900, these rhinos are deemed to be critically endangered, due to poaching and their poor reproductive cycle. It is important for this animal to succeed is to get a better understanding of the reproduction cycle. In clinical samples, blood or urine samples are routinely taken, this is not feasible with rhinos. Hence faecal material is taken, with the initial study looking at progesterone cycles. By monitoring the progesterone levels, it was found that the cycles were not normal and after further investigation it was found that the animals were too heavy. Monitoring of the progesterone also allowed better timing for the keepers to ensure that the animals stood the best chance of a successful mating. The presentation then moved on to how to analyse samples remotely as opposed to monitoring samples in the controlled environment of the zoo.
A swap in the presenter saw Becky describing the issues of trying to collect samples in the plains of Kenya. Becky, known locally as the pooh queen, highlighted the issue of not having a reliable electrical supply. Trying to store the samples in solvent also poses real problems due to sample degradation and evaporation. The solution to all of these issues was to use the SPE at the site of sample collection, and then have the sample stored onto the cartridge. Becky went through the details of the assay which was targeted towards capturing a series of polar metabolites. She stated that although the assay was not as good as the lab assay it performed well and there was a direct correlation, albeit at a lower recovery rate. One of the key criteria of the assays was to be simple and not to be reliant of a constant electrical supply. Becky then described the figures of merit, with a strong correlation for Zebras and elephants existing between the SPE methodology and immunoassay approach. She then started to focus on the future, stating that the zoo was keen to get a better understanding of the metabolism to help the breeding pair become more effective.
Elizabeth Crawley (Elanco)
The bio-analytics of drug development in human health versus animal health
Elanco is a company that does animal wealth fare, and Liz focusses on drug design for animal health. Liz described the structure of Elanco which has a focus on animals that are being looked after, whether that be companion animals or animals bred for the food industry. Liz went through a comparison between animal healthcare product development and pharmaceutical development. She stated that human drug development typically takes 10 – 15 years, however for animal health it is about speed of delivery to the market, with typical drug development times being reduced to 2-5 years and the cost being 1/10th of that spent on human drug development. One of the advantages of the animal drug development is that there is no preclinical stage of development, the research goes straight into the animal under investigation. However since animals do not talk, the issues that are being addressed tend to be much more visible than with diseases associated with humans.
Liz stated the work is regulated by the FDA in a similar manner to human drug development with the regulatory authority for animal wealthfare being the CVM (Centre Veterinary Medicine) in the USA and EMA in Europe. In humans it is necessary to do metabolism ID, PK, ADME, carcinogenicity and toxicology (LD50), initially in animals then moving to clinical. With animal drug development it is allowed to go directly to the animal patient, although it should be stated this is not the norm. In terms of requirements there is an onus on the manufacturer to ensure that the drug is safe. Liz did state that pain drugs for animals were very difficult to develop for obvious reasons, as there is minimal feedback from the animal patients. It should also be noted that for drugs that could end up in the food chain, there are tests performed to ensure that the food derived product is safe for human consumption.
Liz then discussed what a typical submission, NADA (New Animal Drug Application), required, which is comparable to the requirements set out by the FDA for human wealthfare.
In terms of bioanalysis the following are required;
• Animal safety
• Chemistry manufacturing and control
• Environmental impact
• Human food safety, but not for animals which will not be eaten.
It was noted that animal health is a decade behind human health in terms of regulatory aspects. European guidelines are generally stricter than in the US, and as a consequence standards are set by the EU guidelines.
Liz then moved to discussing the future of animal health, stating that there are a lot of developments that will come from human health drug development. Areas where she felt there would be some developments are;
• Addressing issue of increasing numbers of livestock
• Incorporation of diagnostic and genomics
• Aging issues associated with companion animals
• Biomarkers and PD
• Species specific drugs
Howard Parker (Alderley Analytical)