Work experience week at CPI

Ribbon structure of an Adeno-associated Virus (AAV) 

After a relaxing start to my summer holiday I arrived at CPI's Biologics Centre in Darlington on Monday 1st August 2016 for my work experience week on the train from Durham which was only a 15 minute train journey. I was given a card to swipe for all of the locked doors, and set up on the site's computer system with a CPI Email. Jonathan Gunnel, who I had emailed previously to set up the week, was my supervisor who arranged the things I would be doing. He graduated from Leeds University with a BSc in Biochemistry, so had a good idea of the sorts of practicals which would come in useful to have experience of for my university course. He showed me a quick tour so I knew where everything was. A short while after I was given the full safety briefing and hazard talk, and by 11am I had been invited to come to the fortnightly meeting where I was shown the details of the stages of projects and I than to see how the financial side of CPI worked. I was then set up with a desk and computer with an account to email and I got a full sense of just how integral teamwork is to CPI; the office space I was in was on a floor with around 50 other people and the seating was arranged so that all of the downstream scientists and upstream scientists were in the same area, but just across from them were the marketing team, and behind them were the financial team, admin, programme writers, everyone all able to communicate easily and face to face if need be. This sort of team work is needed for a place like CPI, to complete projects there needs to be ease of communication for the smoothness of progress. 

Centre for Process Innovation's National Biologics Manufacturing Centre 
Darlington 

On the scientist side of CPI there were three main departments; Upstream, Downstream and Analytical. Simply put, upstream cultures cells to produce specific molecules or proteins, downstream get given the "soup" of everything those cells have produced and are tasked with extraction and purification of a single type of protein and Analytical then test rigorously to make sure that the purity and correct protein or molecule has been collected. In my week I spent time with al three departments, but the main area of the work I was doing was downstream. 

Centre for Process Innovation's National Biologics 
Manufacturing Centre Darlington 
An industrial scale ion exchange chromatography column

For the first task, I was asked to research the basics of protein separation: Ion Exchange Chromatography, size exclusion, hydrophobic interaction and reverse phase chromatography. After a bit of searching around on the Internet and looking in some biochemistry books, I (hopefully) understood it: for ion exchange chromatography, a column is packed with beads made of, for example, cellulose with a negative carboxylate side chain and therefore negative charge overall. All proteins are made of a series of amino acids which each have a specific charge, hydrophobic affiliation and pH when in solution. These factors mean that every protein is slightly different overall, and by modifying the conditions of the beads and solution you can find a point at which the proteins which are highly positively charged all get attracted to the beads in adsobtion bands. The more positive the protein, the higher the band will be, as gravity or pressure pushes the solution down through the column. If you're trying to separate an overall negatively charged protein from the positively charged proteins, this method should allow the positively charged proteins to be repelled by the beads and so pure negatively charged proteins are collected at the bottom of the Ion Exchange Chromatography column. Each protein has an 'isoelectric point' (due to the addition of many amino acids with different charges) at which there is no overall charge on the protein. By controlling a buffer solution you have control of the rate at which the ions exchange. The beads can then be washed in a stage called Elution where a saline solution can be pushed through the beads, expelling the positively charged proteins to the bottom of the column. The Beads can then be used again, which is a good thing, because I was shown industrial scale columns for which which the beads are specific to the process and can be extremely expensive.

Centre for Process Innovation's National Biologics
 Manufacturing Centre Darlington
Gold Nanoparticle solution.. different to what I expected. 

Centre for Process Innovation's National
Biologics Manufacturing Centre Darlington 
Centrifuge make sure to balance it out!

Part of sitting in an open workspace means that everyone else is working near you on different projects, and I became friendly with a couple of other downstream Scientists who were working on a project which involved insulin attachment to Gold nanoparticles! This was the sort of thing that they give you in a Biology textbook for a 'did you know' box, but I was allowed to watch the process to understand what they were doing. I was surprised the Gold nanoparticle (GNP) solution was a dark dark brown-almost black- not dissimilar to Pepsi or Coke in colour. You just never think of metals as great absorbers of light, especially gold but at the nano scale it's not the same world as we are used to on this comparatively giant scale. Their task was to find the most efficient concentrations and ratios of GNPs to water to insulin for the drug's delivery in the human body, perhaps so that less insulin needed to be used because it could be more localised using this method or had more bioavailability in the human body. This was the first time that I saw the JANUS liquid Handler or 'the robot' as they all called it. Later in the week I programmed and used it, which was good to see in comparison to manual pipetteing.

Centre for Process Innovation's National
Biologics Manufacturing Centre Darlington
JANUS Liquid handler 

Their method was to use a fixed concentration of insulin and to dilute it with the water and GNP solution in different ratios in a plate with 96 wells so you can carry out 96 different versions of need be (or 32 and repeat each reaction 3 times) then Centifuged at 3220x Gravity for 5 minutes, so a pellet of
(hopefully) GNPs bound to insulin were collected at the bottom and then the robotic liquid handler removed the superb start without disturbing the Pellet at the bottom. They then conducted a BCA protein Assay (bicinchoninic acid assay) was used on the supernatant to assess how much of the insulin was left to then work out how much was bound to the GNPs on the pellet at the bottom of the well. The BCA assay turned green- apparently due to Cu²⁺ and ³⁺ ions- if not much protein was present and a deep purple if there was plenty of protein- you can quantitatively use this data if you enter the plate into a colourimeter and use 562nm green light. They explained to me the importance of balancing out the centrifuge, because if you don't it will apparently begin hopping around the room at high speeds! As shown by the image, some results with hardly any protein in the solution were obtained, so were successful. 

Centre for Process Innovation's National
 Biologics Manufacturing Centre Darlington 
Bicinchoninic Acid assay (BCA) 1 min after assay

Centre for Process Innovation's National
Biologics Manufacturing Centre Darlington 
End result of the BCA assay

Reverse osmosis seen through a scanning electron microscope

In the next couple of days I was asked to do some research around the different types of filtration that downstream scientists use to filter out impurities or to leave only larger molecules behind, either way separating them from other unwanted products of the cells. micro filtration, ultrafiltration, nano filtration and reverse osmosis are all different types of scientific filtrations which can be performed on solutions, with varying results. Micro filtration (0.1 micrometre pore size) is the usual type of filter paper which can be bought snd will filter out bacterial cells, whole cells and any suspended solids, such as the gold nanoparticles mentioned above, because they are around 300 nanometres across ( = 0.3 micrometers) however it does not filter out any ions in the water or virus cells. Nano filtration uses pore sizes of around 0.001 micrometers and so can take large ions out of the water such as calcium ions, meaning water can be de-hardened. The most extreme form of filtration is called reverse osmosis because it will only allow water to be let through, and salt ions will be flushed out, desalinating water. Sounds like a great solution to clean drinking water but unfortunately it requires a lot of energy to move the water around the filtration tube and isn't easy to do.

 I was introduced to another method of Protein assay, 280 nanometer Absorbance. To demonstrate this, one of the downstream scientists and I made up different concentrations of BSA  (Bovine serum Albumin- aka Fraction V) from crystalline powdered, pure protein with water.  This test is comprised of a small machine that looks like a miniature George Foreman grill, with tiny metal studs with pores of lights on the base of it, onto which a fraction of a microlitre of the Bovine Albumin serum is pipetted. In every protein there will be some amino acids with an aromatic side chain (such as proline, tyrosine or tryptophan) and these aromatic side chains happen to be very good at absorbing Ultraviolet light at a particular peak of 280nm, so the UV light is shone from the piece of equipment and can be observed in different intensities as it shines through the droplet and is detected on the other side when the lid is closed. Some computer programmes can map this out and tell you the exact concentration of proteins left the drop. The only drawback this method has is that the entire solution needs to be homogenised before pipetting onto the 280nm machine because otherwise you may get a particularly concentrated or weak part of the solution. There is also a coefficient we had to divide our results by which was specific to BSA, but we found that our results were spot on the the concentrations we theoretically had made with a percentage difference of around 5%. The limitations of 280nm absorbance are that it is very sensitive to non-protein matter, for example cell organelles will absorb almost all of the UV light, or even just a mixture of proteins since each has their own specific coefficients (which I assume depends on the quantity of aromatic rings present in the protein structure?).

Protein model of one of the serotypes of AAV

Every week or so at CPI they have a presentation  made by one of the members of staff on a topic or project that they have made progress with recently. I attended the week's lecture by Dr. Cristina Matos on the development of an industrial manufacturing platform for Adeno Associated Virus production (AAV). AAV is a very small virus- only about 20 nanometers in diameter- and lacks a protective lipid coating that most viruses have. There are 12 serotypes (types of virus with distinguishably different surface proteins) and although AAV is not harmful usually, particular serotypes are more effective than others at infecting certain tissues in the body. For example AAV8 is particularly suited to gaining access to liver, heart is suited and AAV1 6 and AAV7 is for skeletal muscle cells. I did not understand much of what Dr. Matos was saying because it was so in depth and she was presenting to a room full of scientists, but I enjoyed it nonetheless.

Next the Analytical department took me on a quick tour of their domain. I never knew there were so many different types of mass spectroscopy- gas chromatography (GCMS), Liquid chromatography (LCMS), and high performance- HPLCMS, even CEMS- capillary electrophoresis. I had not heard of electrophoresis before I saw this machine, so I have done a bit of research around it as I'm sure it will be something which I will be doing at some point in the future. Electrophoresis is defined as a way to separate macromolecules by putting an electric field through a gel. It can be used on proteins, RNA

and DNA, and usually involves a small gel filled chamber with wires attached at either end to create a difference in charge between each side of the chamber, pulling the molecules towards one end at different rates. the faster the rate the further it gets towards the positive terminal and so you can determine the approximate mass of the molecule band seen. Another curiosity I found when researching this type of analytical technique was the unit of mass used, the Dalton (Da). 1 Da is equal to 1 Au or 1/12th the mass of C-12, so the mass of Hydrogen is 1.0007 Da and carbon is 12 Da. Most proteins and DNA are measured in kDa (kiloDaltons). Titin, one of the biggest proteins ever found is 3.86 million Daltons, and around 30,000 amino acids long. so this means the rough mass of an average amino acid in this protein is about 128 Da- a useful conversion if you know the mass of any protein: Mass of protein in Da divided by 128 = approximate number of amino acids. (assuming that all amino acids are in similar ratios to how they are found in titin)

Centre for Process Innovation's National
Biologics Manufacturing Centre Darlington 
Me, all kitted up and pipetting into Eppendorf tubes

One of my favourite individual sections of practical lab work I was introduced to was the immunoassays. In Biology at school we learn how antibodies are Y shaped proteins which can sort of grab onto specific antigens of cells and because they are so specific they can be used by the human body to identify foreign objects or flag up harmful proteins or cells for destruction in the immune system. The Enzyme Linked immunosorbent assay or ELISA as it is commonly known is something which blew my mind when looking into the workings of it. You attach a selection of antibodies to an enzyme which takes colourless substrates and creates a coloured product, these antibodies are then introduced to a solution which you are trying to find a protein in, and they bind to this protein. You then wash away any other antibodies which are not attached to their corresponding antigens and add the two substrates to the solution, which should be catalysed by the enzyme into the coloured product. I was also introduced to the concepts of MIA and RIA- Magnetic immunoassay and Radio immunoassay. The basic idea with these is that you attach an antigen to a molecule of interest (if it does not already have one) and then you label a corresponding antibody with radioactive isotopes of carbon for RIA, or for MIA you attach a magnetic group onto the antibody which can then be detected extremely precisely.

Centre for Process Innovation's National Biologics 
Manufacturing Centre Darlington 
The JANUS pipetting machine uses syringe-like tubes to
create negative pressure and draw solutions up into the heads.

For the last day of my work experience I spent the morning logging new pieces of equipment into the warehouse's computer, and created a safety form for each of them. every piece of equipment from the tiniest 2ml eppendorf tube to a 100L bioreactor must be tested by a member of staff after one of the forms is made and only then can it be used for any new processes that need to be completed. There had been complaints from another member of staff that one of the liquid handling machines had not been accurate enough in pipetting when using a certain type of tip-(different tips can be programmed to be attached to the machine in order to minimise contamination or separate out two layers of a solution). I worked with another downstream scientist to create a protocol for the liquid handling machine to follow into order to test the exact accuracy of the machine using 2 different tips against manual pipetting. The first step was to prepare a batch each of BSA through manual pipetting. The only way to reliably create small solution concentrations by hand is through serial pipetting so I worked out the concentrations and ratios that would produce them, down to a 0.02 ug/ml and then produced them from the same crystalline BSA used before. After we had both made up 90 eppendorfs of various concentrations via serial pipetting, we set the liquid handler to work on its first task; creating the same numbers of Eppendorfs of the same solutions on a 96 well plate with just the normal head (i.e. no tip). The time difference from us creating ours to the robot doing it's was staggering. We took around 45 minutes, but the robot was finished in 5 due to having simultaneous capabilities of pipetting whole rows- (each head had to be washed in a water bath after each cycle to prevent contamination). The next set of 90 Eppendorfs was completed by the robot again, but this time was done using the conductive disposable tips, meaning that they could be sent an electrical signal and could stop pipetting as the machine reached a solid layer at the bottom. more importantly for our experiment was the disposable part of their name, so there would be no possibility of contamination because after every single solution was made, they were dropped off into a chute connected to a bin. This again ran for a very short time and the 90 Eppendorf tubes were collected.

Centre for Process
Innovation's National Biologics
 Manufacturing Centre Darlington
 JANUS robotic liquid handler

We used the 280nm absorbance machine mentioned earlier to quantify the concentrations of BSA we had made. I was then tasked with taking the data and preparing a spreadsheet with accompanying graph to work out the source, if any, of inaccuracy that the data reported and present my findings to the rest of the downstream department. I logged all of the 90 Eppendorf measurements in from the 280nm absorbance machine and averaged all of the readings. It was very pleasing to see that my own pipetting skills were a fraction higher or lower than the other Downstream scientist's who I had been working with, so we knew the were accurate baselines to use and compare the other two lines against. As I added more and more data to excel, the emerging graph showed that the single heads were at a slightly higher concentrations than our solutions, but only by a negligible amount at the lowest of concentrations- probably through small amounts not being entirely washed off. The real source of inaccuracy was found when I began to log in the disposable tips- each one at a far lower concentration than previously expected, and the graph showed a lack of linearity to the results obtained too; where the previous results were all almost a straight line, this disposable tip line had fluctuation all through the results. During my small presentation of my findings to the other scientists I showed this, and then suggested the cause of the error to be systematic, as I had noticed small bubbles of protein solution sometimes left on the tips after they were supposed to have expelled all of the solution. This would correspond to a random error in the concentrations, but would always make it lower than the original three lines, as was shown on the graph I produced. We had located the source of inaccuracy!

Centre for Process Innovation's National
Biologics Manufacturing Centre Darlington
Programming the liquid handler to carry
 out a set of instructions for dilution 

The suggestion after I showed my findings was that an air blow step was to be added into the instruction commands for the robot when using the disposable tips, in which a small amount of air would be blast through the end of the tip to pop any bubble left and add to the concentration.








Through my entire week at CPI I learned so many new skills and ideas beyond those taught at school which will no doubt be incredibly useful as I continue my scientific studies at university. I can only thank them for giving me the opportunity to do this. It was a great week and CPI is definitely somewhere I could see myself being employed in the future...

Wellcome collection visit, London

On the 14th of July I made an impulsive London trip, because I was bored in the holidays. I didn't really have much of a plan, but I knew I wanted to go to the Wellcome foundation to have a look around at their exhibits which are always interesting and maybe have a peek at the universities (ICL and UCL) as I like the idea of student life in London.

After arriving on a delayed (as usual) train to St. Pancras station I wandered down the Main Street towards the welcome collection, stopping hallways in the British Library, where they had a great exhibition with many priceless books that have contributed to the foundations of both England and the world. Things like the Magna Carta (Which when translated from Latin means big charter- sounds much less important!) original Shakesperes, Lindisfarne Gospels and the oldest recorded bible were sitting in cases under dim lights so as not to fade the inks and papers. 

the bookcase of life

I carried on down to the Wellcome collection and had lunch in the cafe there before looking at the upstairs exhibit as I knew it would be much more to my interests, a medical exhibition and an exhibition on the states of consciousness. The first thing you come to in the medical exhibit is one of my favourite things I saw that day; a whole bookshelf filled with about 120 books, each one being a specific chromosome  of our 23, and they all contained the genetic code of ATCG written in tiny font. That bookshelf is what makes each one of us unique. In those books are the literal instructions to make a human, only 3,000,000,000 steps to follow.. Yet all of our cells contain this vast expanse of storage and can multiple and carry out more of these instructions than we could ever hope to through the wonders of biochemistry. Another thing that got me thinking was the pure weight of the information in those books, yet a nucleus of a cell is probably billions of times less massive, I wonder how much it would weigh to store it on a computer of today's standards? Would man or machine win? (I did some calculation but got rather confused with the numbering systems of bits on computers and so the answer turns out to be around 200Gb, which is about all the memory of a small laptop, I doubt you could fit that into a cell) this further prompts me to wonder if we will ever use cells as a form of data storage?( I think) They're more compact memory storage than computers, but the drawbacks would obviously be things like mutations and base deletion, substitution etc. Maybe if we could engineer a binary cell? Instead of ACGT only have 2 "nucleotides" and read them off like numbers 1 and 0 in bits... Anyway, I'm digressing. 

Chromosomal pairs of socks

There were some of the Gunther Von Hagen plastinates slices there too (pictures at the end of the post for those who aren't squeamish) which are always amazing to see, tracing out the individual pathways and seeing how all the tissues are interconnected is fascinating- I have been to three body worlds exhibits in the past and I plan to go to one in Newcastle next month too- all of his work is quite thought provoking, bridging the gap between science and art. Some more art/science installations included glass blocks with resin shapes highlighting areas of the brain which showed increased activity when participants were given things to see touch taste smell and hear. Our brains must be very active if we are doing all five things at once. And a really funny piece was sock chromosomes! The artist had made the typical chromosomal lineup out of pairs of socks, which was very amusing. 

The sight 'portion' of the brain

Moving on to the consciousness exhibit, I already have quite a lot of knowledge about consciousness states and sleep because of the work my mum does. When I was about 12 she gave me a book, 'the man who mistook his wife for a hat' which blew me away. She was a neurophysiotherapist and now works in case management of people with severe brain injuries, so she has explained to me the sorts of states of consciousness you can be in. Coma, Vegetative state, MCS (minimally conscious state) and emerged, then past that there is a broad range of neurological problems people can have from disinhibition to memory problems and emotional disconnection and also connected physical problems such as paralysis or muscle spasticity. I understand a lot of these more neurological concepts because of the work she has done and the amount I am involved in through administration in her case management company, hopefully this knowledge will come in useful in the future whatever I decide to do. Neuroscience is such an interesting topic!

Original Santiago Ramón y Cajal drawing of brain cells in exhibit

I saw the original drawings of the grandfather of neuroscience- Santiago Ramón y Cajal, which were so intricate and beautifully drawn. He himself was an artist, but was persuaded into medicine by his father and therein his passions combined and he is probably the most noted neuroscientists of all time. (I wasn't supposed to take pictures but it was too good of an opportunity not to!) 

I had a go at some Synesthesia training, synaesthesia being a neurological condition in which sensory neurones are mixed up causing odd senses to be used, for example in extreme cases people report particular smells when they see a certain colour or maybe a taste when they hear a sound. 

I was given 5 minutes to associate colours with letters, and then a quick fire round of linking the colours I had seen with letters and vice versa for another 5 minutes until I knew them inside out, (a was red, b was green, i was orange, etc) 

Then I was given a passage to read about synaesthesia in black and white, but each time I finished a paragraph, every "e" or would be replaced with a purple block, covering the letter, then every "i" being replaced with an orange block and still I could read the words perfectly. However I thought of this not really as synaesthesia, but more as a "full in the blanks" where if you muddle the midldle of wrods you can stlil readthe setncnes pefrctely well beaucse yuor brian fills in the gaps (Case in point) but as it got so that more and more letters were filled as blocks of colour, even full words were just series of colours... And I could still read the colours perfectly!!! What an incredible machine the brain is.

The last section was a section on sleep, and how sleep problems can be dangerous at times. Instances such as sexomnia have been documented and reported, with scientific backup and have even exonerated people from prison for crimes like rape and sexual abuse because the person was not in full control of their actions. Violent attacks and even murders have been perpetrated while asleep as they showed a video of a man plagued with violent sleep patterns in his REM (rapid eye movement) sleep as he attacked the dummy body left in the bed next to him with disturbing ferocity. This is often a result of insufficient compensation by the brain, as it paralyses your body while you sleep (except the heart, lungs and small limb movements) but if it doesn't paralyse you fully, you can "act out your dreams". The opposite of this is when the brain Kees the body paralysed after they have woken up, and is a common experience for some. They feel that they are rigid in bed and although they can open their eyes, they are entirely unable to move or speak. This only usually lasts for a few seconds but is scary to experience I am sure.

I then wandered downstairs for the new exhibit on the ground floor. It was entitled 'This is a Voice' the first room I walked into was very eerie- there was nothing in it except it had spiked foam sound absorption tiles on every wall and the ceiling, meaning that when you clapped or whistled the sound was immediately dampened and still. You could hear your own breathing and heart rate better than the voices from the next room. It felt like someone was holding a piece of cotton wool over each ear.

The next room focused on the origins of the human voice, how through the process of evolution our voices have adapted and changed to suit our needs. From earlier neandethaals characteristic 'grunts' to the pop stars vocal acrobatics of modern day, the voice of the human race is a very flexible thing. Human voice originally evolved for the purpose of song and social bonding rather than information exchange. Early hominin groups became larger and physical grooming was no longer efficient form of bonding, so the voice kept individuals emotionally connected creating alliances and group dynamics. Reformulations to the larynx and bipedalism with changes in facial anatomy helped to further the human race's rate of evolution. There are significant advantages to being able to have a strong voice, to warn off predators or other tribes, and increase group cohesion among others. 

I was not as taken by the next room of the exhibit: a man in a suit was stood in front of a group of an audience and he had a sheet of music he had written himself, as he explained that he would demonstrate the variety of the human vocal cords. I expected him to perhaps sing some opera and hen maybe falsetto or perhaps even beatbox? I was greatly mistaken.. He exploded into high shrieks and low bellows amidst whimpering and full shouting, none of it with actual words behind, only sounds. 

I found it extremely hard to keep a straight face. This smartly dressed man bawling like a baby then whooping like a Native American war cry then screaming.. And all of the people watching were deeply interested with passionate serious looks on their faces which only made me want to laugh even more. I exited quickly and politely.

University College London is directly behind the Wellcome collection so I went and grabbed some undergraduate prospectuses from the office there and turned straight to the biochemistry and natural sciences courses.. I feel like they're the courses for me! 

Had a great rest of the day, went to Knightsbridge for a look around harrods and all the posh shops, then over to embankment, you can't go to London in the summer and not visit The Houses of Parliament and Big Ben. 

Visit to CPI

A few weeks after talking to George Ewart as mentioned in my last post, he contacted me to say that he had been at lunch with an old friend who was a non executive member on a board of a company called CPI (Centre for Process & Innovation). At this lunch he mentioned the talk that he and I had and his friend offered to take us both on a tour of CPI, to show us around the centre's facilities and meet some of the members of staff. Of course, this was fantastic news as this could be a great example of a place that I could see myself working at in the future, and it would give me an opportunity to understand more of the business and see how it drives scientific processes in the real world. 

I got in touch with Sandy Anderson, the non exec board member, who was kind enough to arrange a full day's programme for George and I, as he had not seen the centre before either.  

I decided to do some background research on the centre before I went, to understand more about what I would see. CPI employs just under 400 people, and are based in the North East of England, with their main site being in Wilton. They were set up by ONE North East, and their current CEO was Nigel Perry. They particularly specialise in working with other firms and companies to develop commercial products, medicines/biological testing and technologies. I also researched around a few of the people I knew I would be meeting, such as Sandy Anderson OBE who was actually a governor at my school for almost a decade, and stopped when I was in year 8, in 2012. Alongside his current position in CPI, he has also been the chair and pro chancellor of Teeside University as well as chair of various other companies including senior vice president at ICI. Dr. Lucy Foley, the head of Biologics centre in Darlington (which I would be visiting later) who studied chemical engineering at UCL, and went on to become a teaching Fellow at Newcastle University, and Dr. Tony Jackson who had previously worked at ICI, and will have no doubt shared connections with George. He then worked under AzkoNobel for 6 years, being project director and research team leader before coming to CPI and being their head of Research at the Formulations Centre, which George and I would also be visiting later in the day. CPI had recently gained £15 million of funding from a scientific council for future formulation projects, set up two new buildings in the NETPark in Sedgefield, including a graphene research centre- something which my chemistry teacher would absolutely love to see. 

On 19th of May, I caught the train to Darlington and was picked up by George as we made our way to the Biologics centre to meet Dr. Lucy Foley. I had not realised that this centre was so brand new! Everything was beautifully designed in this centre, with some very cool panoramic views around the main conference rooms of the labs. We were given a full floor-to-floor tour of the place, from the liquid nitrogen tanks outside to the heart of the machinery and computer modelling rooms. They even had a feature that allowed an iPad to be pointed towards any wall and show-in real time- the pipes behind the walls or ceilings to ease the problem of fixing pipes or doing maintainence accurately and efficiently. We had a talk with Dr Foley in which she explained all of what goes on in this centre, and showed us particularly her area of expertise, fermentation. They had around 50 small (maybe 500ml?) fermentation reaction vessels in one room which were being constantly measured and had different amounts of reactants in them, all hooked up to a self producing graph to find the optimal mixture of ingredients and conditions so that they could then scale up to the 10 or even 100 litre vessels in order to mass produce the product. In the one and a half hours we had there, I was very impressed by the whole place and asked Dr. Foley if they would be interested in linking with my school, for the possibility of a school trip or maybe for a lecture to one of our science societies as we have such a variety of good chemists and biologists at Durham school, to which she said she would put me in touch with their STEM co-ordinater to arrange this. She also offered me a week's work experience at CPI during the summer which I happily accepted! 

The next place we went was CPI Wilton; where the company was based. Upon arrival we had lunch with the CEO, Nigel Perry. He really helped me to understand the business side of what CPI do, and explained in more detail the projects that were taking place all around the area. The lead scientist there also showed us around the labs that they had, with all sorts of amazing equipment. Some of the coolest things I remember were polypeptide sorting machines, they would be moved parallel across a membrane with specific conditions on either side to facilitate the movement of certain proteins and not others. By passing them parallel to the membrane you do not change their structure by force and by altering the conditions (for things like specific amino acid charges and polymer shape) you can vary the range of polypeptides which cross, selecting them, in effect. I also remember a fluid which the polypeptides would travel at different speeds through and this was hooked to a computer which would be able to identify the identities of the proteins collected. 

The final place we went on our tour of CPI (which was turning into a tour of the north east of England!) was over in Sedgefield, at the "NETPark" where I met Dr. Tony Jackson and had a look into some printable electronics and Graphene research they were doing in their brand new centre. He explained how they were revolutionising the precision with which they could print and even said they could now print at an atomic level, around (1x10⁻⁹ metres) and showed us some incredible examples of technology which had been printed and woven into clothing and touch pads which were printed into a seemingly solid table surface which were unbelievably sensitive and gave a beautiful image on a computer screen. The graphene was interesting especially to talk about because it was a section of the GCSE chemistry course and we had to learn about it as a giant covalent structure. One thing he did mention was graphene's possible asbestos-like properties, as far as being unreactive enough to slice DNA apart and cause mutations, and so as a safety precaution until more is known it unfortunately has to be kept behind glass or in containers! He also talked to me a lot about his particular route through Both CPI and AzkoNobel and his experiences of leadership in interesting foreign countries including Germany and China as well as how his family life had affected his career path and his need to balance the two. This gave a deeper understanding for my future and will be something I am sure I will need to take into account at some point.

I loved the day I had at CPI, and I hope to do many more similar things to it in the future. I am so thankful to George for introducing me, and then to the whole of the staff who showed us around at CPI and for organising such an opportunity for me. Defintiely somewhere I could see myself working in the future... 

EDIT: Work experience commences in the 1st wee
k of August and a school trip will be arranged hopefully in Sep/Oct this year. 

Body Worlds exhibition Newcastle

A few months ago I noticed that the centre for life (Newcastle's science centre) was hosting another Body Worlds Exhibition, similar to ones I had previously seen in Amsterdam and a previous Newcastle exhibition. I loved these exhibitions as they were so interesting to me and like the Wellcome collection in my last post, blurred the lines between art and science. In fact I used a lot of Body Worlds' exhibits for inspiration for my GCSE art module on structure, I did the structure of the anatomy of the human body.

If you haven't seen Body Worlds before, I'll explain a bit about it: The brainchild of Dr. Gunther Von Hagens, an Austrian doctor of medicine, who wanted to educate and show the public the wonders of human anatomy, Body Worlds is a travelling exhibition of real human specimens who have been "plastinated" in a chemical process which removed the body tissue and replaces it with an exact plastic replica. The result is an awe inspiring detailed 3 dimensional model of how the human body fits together and what you would really look like under the skin. It's sort of like a human body cross section book, but in real life and much more detailed. Over 40 million people have viewed a Body Worlds exhibition, and it has been a massive success with exhibitions in over 100 cities worldwide from 1995 to the present. Although some of the images you are seeing around this post may not be to your liking, I highly recommend going to experience any exhibition you can find near you. 

Both of the pervious exhibitions I had seen were about the human body, one being Pulse and the other Vital, so they particularly focused on the circulatory system and internal organs respectively, however this exhibition was entitled "Animal Inside Out" and featured all sorts of organisms from the kingdom animalia, including things from pigeons and frogs to a giraffe and an elephant! I encouraged a couple friends who were also interested to come with me, even though one was actually a vegetarian! The variety of organisms was one for he things I enjoyed most about this exhibition as they differed from the same human forms (normally you aren't allowed to take photographs of the plastinates to respect protect the identities of the plastinated human's, but as this was animals photography was allowed so I took many pictures to post on the blog) 

I have included a section which I found interesting at the very beginning of the exhibition on the process of plastination, invented by Dr. Von Hagens himself, using Acetone to dissolve the soluble fats and then replacing the structures with Silicon which enters the cells through a vacuum pump at which point they can position the limbs and tissues how they need and will then harden the body for display. 

The first plastinate we saw was of a Dog, posed catching a frisbee, jumping in midair. The muscle groups which you can see are particularly accentuated are the neck, shoulder and jaw muscles and this makes sense as the bulk of a dog's power is from its back and shoulders as it had the evolutionary advantage of using its mouth to bite and neck to balance during running (such as is particularly noticeable in a greyhound running at full speed) the red groups are the muscle bulk and the white parts are tendons, connective tissues and joints, such as is visible on the front paws and wrists. 

The next three exhibits were great to see in a row, three ostriches showing the different layers of muscle bone and circulatory system that was present all over the body. Comparing all of these techniques made me appreciate just how  much of a complex organism like an Ostrich is. Muscles interwoven with bone and blood vessels branching out through the muscles in a seemingly arbitrary pattern, but maintaining constant blood flow to all aerobic reactions needing to take place in the ostrich's muscles. The genes and hormones that regulate these processes of where things grow and defining boundaries between tissue types must be so complex, no wonder there must be so much information in every nucleus of the cell. 

I find it fascinating to take my knowledge of biological systems and how they have helped shape the physiology of the organisms in the exhibition and then think deeper about the millions of billions of biochemical processes that would have to have taken place to make all of this possible, and then even further down to think about how the laws of physics and characteristics of chemicals and elements have shaped the world we live in, and how if they had been at all different would I still be writing this as I am now?







One of my favourite plastination techniques Von Hagen uses is the plastination of capillaries, as it makes you realise just how dense your network of blood vessels is, that if all the capillaries are used, there is a surface just as the skin would be covering the entire shape of the horse. No cell in the body is less than 0.1 micrometers from a capillary. you may as well be looking at the horse's head actually dyed red, but in fact if you were to use a microscope you would see that every single capillary is separate and gaps are left for cells. The Ostrich Circulatory system mentioned and shown above was only showing the major blood vessels, as if it has been done like the horse you would only see a red ostrich.






The next two exhibits were two of the most impressive: a Brown Bear and a Gorilla, both gigantic and imposing in stature with their huge muscle groups on their shoulders and torso, evolved and adapted to be high ranking in their food chains over millions of years. one of the only Human palatinates was featured in this exhibit next to the Gorilla, to show how closely related we were but also how different our physical bodies are. Unfortunately photography of the human palatinate is not allowed, but i was able to take a picture of the human brain, and the difference between the size of the gorilla's and the human's is what was extraordinary, the human's was not much bigger, yet we are the only species capable of language and communication. I suppose it was slightly misleading as the usual way the comparison is made is by brain size in relation body weight, but it made an impact.

The biggest two plastinations I have ever seen were left until last: an elephant and a pair of giraffes. the giraffe was so tall, and was split directly down the middle to expose the internal organs from behind. the elephant was expanded, with each layer of tissue being slid away from those under it to make it seem even larger than a normal african elephant. This technique also exposed the muscles and parts of bones or arteries which would not normally be shown as they are buried between layers or at least are not seen easily on the surface. The entire brain and nervous system was laid out underneath the elephant at it's feet, immediately made me realise just how much distance a nerve pulse must have to travel to get from the elephant's brain to it's foot and all the way back in a movement- would this time difference make a slight lag? Would we perceive the elephant's foot as moving after a noticeable delay once the elephant had thought to move it's foot? The brain itself was huge, but in comparison to the body size, had nothing on the human brain, of course. i have ridden an asian elephant in Thailand, and I do remember feeling as though they had some intelligent understanding, more than i have ever felt that a horse or cow had. Their trunks have a finger like feeling appendage to the end of it which they pass far above their head in search of bananas they know you have and (their favourite) a giant leek-like vegetable, which they would crunch through on a walk and you could feel the force and noise of the crunches through the vibrations in the back of their necks where you sat.


The second of the pair of Giraffes was a clever way to display an animal; it was slices of plastinated sections of giraffe suspended from the ceiling in the shape of a giraffe, so you could look at the layers and follow specific organ systems or tracks all the way through the body as the lights shone through.


This exhibit was another great success for body worlds in my opinion, and I think that anyone with a a keen interest in animals, humans, physiology, biology or even life in general should go to see this exhibition, I can't recommend it enough.

Teamwork at Sunderland Analytical Chemistry Competition

The 3 highest achieving chemists from our school were selected to take part in an analytical chemistry day at Sunderland University, and I was asked to write up the day's event for the school newspaper. Although we did not win, I believe we did well as we seemed to all get results that matched up with the motives and the culprit in the end.

Marrow murderer at large!

A year 12 team of Anu Krishna, Fraser Gaines and Will Bowles representing the school were invited for a day of a analytical Chemistry at the labs of Sunderland University on Tuesday. They were each given a booklet containing a story of a man called Dave who wanted to grow a prizewinning marrow in his garden; however he had made some enemies in the village and his marrow ha
d subsequently been destroyed by a mystery marrow murderer. 

Three suspects were narrowed down: firstly, Jason, a boy who had his football confiscated by Dave for kicking it into his garden who was believed to have held a grudge and be seen with a large bag of salt near the time of the crime. Next was Ms.Dale, a fertiliser retailer in the village who threatened to sell Dave fertiliser canister that's contents had been replaced with water. Finally was Mrs. Pembroke who had been recently spotted with copious amounts of silver nitrate, a toxic chemical, which she had no conceivable need for and also happened to hold a grudge against Dave and his marrow contest. 

Flame Photometer

Each of these suspects were to be investigated using analytical chemistry methods by each of the three members of the team, with Anu sorting the Silver nitrate scandal, Fraser testing for Phospherous in the Fertiliser and Will working on the salt scheme. 

Will made samples of different concentrations of salt water to be compared with both water obtained from the marrow plant's surrounding soil and soil from elsewhere in the garden. Using flame spectroscopy (where a sample  of a chemical is sent through a methane flame and the energy absorbed is an indication of the proportion of a certain element in the sample) Will found there to be less than 0.01 ppm (part per million) of Na⁺ in the water, and there was no difference between samples taken directly from the marrow soil and elsewhere in the garden, thus exonerating Jason from having committed such a terrible crime.

The Silver Thiocyanate ion titration

Anu was next, using a Ferrous thiocyanate indicator to detect a colour change from milky to grapefruit pink with the thiocyanate ion ([SCN]⁻) when combined with silver nitrate ([Ag]⁺[NO₃]⁻) and 6 molar nitric acid (HNO₃) to form a Silver thiocyanate precipitate when all of the iron had been used in conjunction with the thiocyanate

([Fe]³⁺3[SCN]⁻ with excess [Ag]⁺ means [Ag]⁺[SCN]⁻ silver thiocyanate precipitate is produced.

 By titrating this solution Anu was able to work out there was no significant difference in the amount of AgNO₃ in the soil around the marrow plant, and it was much less than the weakest concentration solution she had prepared in the lab, meaning that Mrs. Pembroke had also not poisoned the prized marrow.

It all came down to Fraser Gaines' phosphate fertiliser activity, as he used the analytical test for phosphate ions; ammonium molybdate-(2[NH₄]⁺[MoO₄]⁻²)

If phosphate was present the solution turned a dark marine blue, and the concentration of phosphate was proportional to the depth of the blue colour. 

Ammonium Molybdate

Fraser mixed varying concentrations of phosphate to use as comparisons, and then tested the sample from the marrow patch and the soil from elsewhere in the garden, finding a distinct difference between the two; the one from the rest of the garden was a dark blue, with a high concentration of phosphate [PO₃]⁻ ions, where the Dave had previously used fertiliser before Ms. Dale had started to be suspected of selling watered down fertiliser, the other from the marrow patch being a light transparent sky blue, indicating a paucity of phosphate ions, which, after using analytical photospectroscopy techniques ( which measure the absorption of light of a substance), Fraser could identify as having the same concentration as the weakest solution he had prepared, showing that Ms. Dale had indeed watered down the fertiliser to sabotage the growth of the prized marrow.

The 'Smoking Gun'

 Having taken longer to complete the analytical side of the practical work, the team was left to write a conclusion with fo
llow up questions, which were answered in a shortage of time and unfortunately did not secure them first place. 

The experience was both interesting and difficult, while improving their understanding of chemistry through analytical practical skills which will no doubt be good experience for university and later life for all three members of the Year 12 team.

Meeting with George Ewart- ICI (Imperial Chemical Industries)

ICI logo- founded in 1926 as the
product of four companies merged.

Through conversation with one of my mother's work contacts about A levels and future interests, it transpired that her father worked in a company called ICI -Imperial Chemical Industries- a chemical company founded in Britain in 1926 and for much of it's history has been the largest manufacturer in Britain. As I am thinking about the future and what types of things you can go on to do with degrees from university in things like chemistry, biochemistry and biomedicine, this seemed like a great opportunity to question what exactly her father did and his general path through life. She told me that he had studied Chemistry at Glasgow University and after he graduated he had been involved with ICI, at first being employed with a more chemistry-centric role to his work, but later rising through the company until he became the Managing Director of ICI's overseas unit in India.
She also explained that he was extremely enthusiastic about encouraging young people into chemistry and related fields, and would be more than happy to have a meeting with me. Of course, this seemed like an amazing opportunity, and so I jumped at the chance to meet Mr. Ewart and ask him about his career and see if he would answer any questions I had about the career path I might want to choose.
Before I met Mr. Ewart I had done some more reading around about ICI and what it specifically is involved in: the company was the result of a merger between four substituent companies; Brunner Mond, Nobel Explosives, united Alkali Co. and British Dyestuffs Corp. In its early days, it was competing with other large chemical firms such as IG Farben and DuPont and it's original base was in Billingham, Stockton-on-Tees, not far at all from where I live. It has contributed to many industrial processes and been key to developing new chemical products, things like:

Pthalocyanin

  Dulux Paint colour pigments (with DuPont in 1932)

  Pthalocyanin(1929), a blue/green dye used in artist's paint and even    used in a large proportion of CDs

  Perspex (1932) and Polyethylene (1937) of course you've heard of these

Terylene polymer

  Terylene (1941) a polymer used   about 30% for bottles and 60%   for fabrics (also known as PET)

  Crimplene (1950) a fabric   material resistant to creases

it also branched out into more Pharmaceutical products, creating ICI pharmaceuticals in 1957 under which things such as Halothane (1951) was produced, an anaesthetic agent. Inderal (1965) a beta blocker, Tamoxifen (1978) was a drug used to combat breast cancer by being an inhibitor of the processes enabling breast cells to grow, therefore it is especially used for men with breast cancer. Between these years it had also been developing many successful Pesticides, such as primiphos-methyl, primicarb, brodifacoum and lambda-cyhalothrin.

 Various changes in ICI's CEOs skyrocketed it's profits and in 1970-75 it was Britain's largest Exporter, and began to make huge business moves such as Acquiring Atlas Chemical industries in America and BNS (British Nylon Spinners) as well as selling other parts of it's company. ICI was included in the FTSE100 while this buying and selling continued until it found itself in debt of £4 billion, at which point it sold its chemical commodities, £1 bn to ICI Australia, and £3 bn to DuPont in 1997. After this, it was taken over in 2006 by AzkoNobel with an initial bit of £7.2 billion, however this was later increased to £8 billion and accepted by ICI. It still has bases in Stockton-on-Tees, Redcar & Cleveland, Manchester, Slough, Ayrshire and Hertfordshire.

Picture of George Ewart
taken for Indian news.

I also managed to find some news from India about Mr. Ewart's division, specifically that the Indian division of ICI was involved in rubber chemicals and explosives, particularly the formation of Nitroglycerine. It employed around 7,100 people at the time.

George and I had a great couple of hours of conversation, as he took me through his career path from his various experiences of great things and memories from Glasgow university where he studied, to his particular memories of his first roles in ICI, where he travelled to many eastern european countries to be the middle man between the chemical producers back in the UK and the sales teams for the consumers in these countries, therefore his role in these positions was to decide on particular prices, as he had knowledge of the costs, processes and yields of these chemical reactions. This gave him confidence in the business sides of things, and he encouraged me to open my mind to travelling to different countries to build experience in foreign places.
He asked me what A levels I was taking and how I was getting on with them. I Replied that I enjoyed both Biology and Chemistry the most out of my four (Bio, Chem, Physics and Latin) and was wondering where in particular he thought might be a good area to focus for the future sorts of fields within the chemistry/biology world to which he replied that he thought biochemistry and biomedicine were probably the most up and coming sectors, due to the newly available technology we are leaning how to utilise. Of course when he was young, chemistry was all the rage, without biology having caught up for example, Crick and Watson only discovered the specific structure of DNA in 1953, and so much more was need to be discovered before the corporations could successfully build business in fields relating more heavily to biochemistry and biomedicine technologies.
Another interesting point raised during our discussion was the idea of being accepted into a company to work, and them sponsoring me through university in order to take me back and have me working in the company with a degree. I have searched around for this sort of opportunity however most companies do not seem to offer this, at least not openly. It was fantastic to have met and had a long and meaningful conversation George, as I hope he will be a useful contact in the future, and continue to be a source of inspiration to me.