Difference between revisions of "Team:UCL/Entrepreneurship2"

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<p class="title" style="text-align:center;">Entrepreneurship</p>
 
  <br>
 
  <br>
 
            <ul class="cos">
 
          <li id="item1">Executive summary</li>
 
            <li id="item2">Product</li>
 
  <li id="item3">Intellectual Property</li>
 
            <li id="item4">Manufacturing Operations</li>
 
            <li id="item5">Business Environment</li>
 
    <li id="item6">Marketing</li>
 
            <li id="item7">Management</li>
 
    <li id="item8">Financial Data</li>
 
   
 
            <!-- 
 
<li id="item9">IPTG-induced protein expression</li>
 
  <li id="item10">Spectrophotometric assays</li>
 
<li id="item11">...</li>
 
                <li id="item12">...</li>
 
                <li id="item13">...</li> -->
 
 
 
            </ul>
 
 
        </div>
 
 
 
<div id="menu3">
 
<p class="title">Protocols</p>
 
            <ul class="cos">
 
              <a href="#summary"><li id="item1">Executive summary</li></a>
 
                <a href="#product"><li id="item2">Product</li></a>
 
              <a href="#IP"><li id="item3">Intellectual Property</li></a>
 
                <a href="#Operations"><li id="item4">Manufacturing Operations</li></a>
 
                <a href="#Marketing"><li id="item5">Business Environment</li></a>
 
                <a href="#Management"><li id="item6">Marketing</li></a>
 
                <a href="#Financialdata"><li id="item7">Management</li></a>
 
                <a href="#Risk"><li id="item8">Financial Data</li></a>
 
              <!-- 
 
<a href="#iptg"><li id="item9">IPTG-induced protein expression</li></a>
 
                <a href="#assays"><li id="item10">Spectrophotometric assays</li></a>
 
<li id="item11">...</li>
 
                <li id="item12">...</li>
 
                <li id="item13">...</li> -->
 
 
 
            </ul>
 
 
        </div>
 
 
<div id="protocolcontent">
 
<div class="default-text"></div>
 
   
 
 
        <div class="about-item1 hide">
 
 
 
          <div class="protcl" id="summary"> <h2>Executive summary</h2>
 
<center><h3> ProKao - we make your microbiome work for you.</h3></center>
 
<p>We are a group of students who became inspired by the theme of their iGEM project and decided to adapt it into a
 
  real world solution. Our products allow users to harness the potential of their microbial flora to help them
 
  achieve their goals and increase their quality of life. The first product we will put on the market is a probiotic
 
  chocolate bar which contains bacterial strains that have been shown to reduce sadness and increase the ability to
 
  cope with stress. This is the first application of what will later become a platform technology that targets a
 
  variety of the user’s needs and goals using different combinations of living organisms and active substances. We
 
  have already filed the first of what will be several patent applications detailing the composition of our product
 
  and the methods required to produce it.</p>
 
 
<p>The market for both probiotics and mental wellness related products is expanding worldwide taking up 38% of the
 
  world's functional foods market. Despite the comforts that come with the abundance of modern life in the developed
 
  world people are far from care-free. We still have the same instincts and brain of pre-historic humans that had to
 
  constantly worry about food, shelter and predators. These instincts manifest in fight-or-flight reactions in
 
  situations where our physical well-being is no longer in question. This chronic stress has been linked with a
 
  myriad of other health problems, from high blood pressure, depression, a weakened immune system and auto-immune
 
  disorders. There are probiotics as well as numerous supplements in the market aimed at improving mental wellbeing,
 
  which is not surprising given the scale of such a concern. By targeting specific groups (stratification) or tailoring
 
  each batch according to a customer’s needs (personalisation) we’re able to leverage a growing body of research to a
 
  greater extent than ever before. Our products will help patients achieve their goals by tapping into the potential
 
  of the most neglected organ in the human body:  the microbial flora, and do so with greater accuracy and control
 
  than any that have come before us.</p>
 
 
<p>Our first product is inspired by our iGEM project: a probiotic that helps people decrease their stress and
 
  anxiety and promote their mental well-being. The bacterial strains we will used have all been shown to produce
 
  beneficial effects in clinical studies and will be combined with other ingredients in such a way as to be tailored
 
  to their personal situation, needs and goals.</p>
 
 
<p>Our stratified range product will be sold at retailers that cater to the health-conscious whereas the personalised
 
  product will be specified by the customer online and made to order based on the information provided. Prokao is a
 
  product meant for daily consumption, especially the personalised version that can be changed progressively if the
 
  customer opts to sample his microbiome on a regular basis.</p>
 
 
<p>Over the past three months we have been advised by academics, consultants, attorneys and several people from the
 
  venture capital fund SOSVentures. Their insights have been invaluable and have motivated us to take this project
 
  beyond iGEM. Our initial funding will come from participating in a business incubator where we will develop our
 
  proof of concept. Within one year we intend to have first product on the market and start working on the next
 
  application. In three years we expect to have developed and patented optimised strains that will be used in our
 
  products as well as have begun to look into developing strains using genetic engineering techniques. In five years
 
  we expect to have our first medical treatment entering clinical trials.</p>
 
 
<p>We intend to start a revolution in microbial based treatment. This technology is in its infancy but has the
 
  potential to overturn the medical establishment in its current form. It will allow for continuous monitoring
 
  of chronic disease to the point of virtual cure. Our ultimate goal is to empower our customers, not only by
 
  reducing their suffering but also to help them achieve their desires.</p>
 
 
</div>
 
 
        </div>
 
     
 
 
<div class="about-item2 hide">
 
         
 
<div id="product" class="protcl"> <h2>Product</h2>
 
  <p> </p>
 
  <h3> Overview </h3>
 
<p>ProKao is a delicious chocolate bar that combines the goodness of chocolate and the benefits of probiotics.
 
  Our chocolate is sourced from organic and fair trade producers, and has been enhanced with billions of
 
  friendly bacteria, which have been shown to interact with your gut-brain axis, to make you happier and healthier,
 
  and help you cope with the stress of everyday life.</p>
 
<p>Once a day you will be able to indulge in our delicious chocolate bar, knowing that not only it will deliver
 
  these healthy bacteria to your gut, but also contains dietary fibers (prebiotics), which stimulate the
 
  proliferation of your natural microflora, without any added calories.</p>
 
 
<img src="https://static.igem.org/mediawiki/2015/2/23/UCL2015product3infographic.jpeg"> </li>
 
 
<h3>How our product came to be</h3>
 
  <p>The idea to create a probiotic to help with stress, anxiety and other mental health-related issues came
 
    from the core theme of our project: the gut-brain axis. Our original idea was to create genetically engineered
 
    probiotic strains but soon we realised that the current legislation doesn’t allow for the commercialisation of
 
    such products aimed at the general public, and only recently have tests with animals began taking place for
 
    pharmaceutical applications.</p>
 
 
 
<h3>A three tier vision</h3>
 
<p>The set of constraints we were initially faced with lead us to pivot away from focusing genetically engineered
 
      probiotics from the start. Instead we opted for the following timeline:</p>
 
  <ol>
 
    <li>A product based on characterised strains and other GRAS (generally recognised as safe) ingredients that have
 
      a track record of efficacy and clinical data to back them up. This will be our minimum viable product,
 
      categorised as a food.</li>
 
    <li>Once the mechanisms behind the effect of different microbiome profiles have been fully identified it will be
 
      possible to create enhanced strains. This can be done without necessarily being subjected to the tight
 
      regulations that restrict genetically engineered products, of course this would limit the scope of our action
 
      to certain techniques (deregulation, generating mutations, directed evolution, CRISPR-cas9). Another option
 
      would be to run our own experiments on particular strains we create or discover and patent them. These products
 
      would also be characterised as foods.</li>
 
    <li>Using synthetic biology methods to enhance the production of particular compounds, introduce the genetic
 
      circuits that we have created to respond to environmental triggers and create ways of controlling the production of particular substances.
 
      This phase offers the greatest potential and the widest variety of applications; bacteria can be engineered
 
      into microbial factories that produce pharmacologically active substances of our choosing and secrete them in
 
      the desired amounts. A product such as this one is beyond the scope of what current legislation allows, and
 
      would only be considered for approval as a therapeutic and therefore be subjected to clinical
 
      trials.</li>
 
  </ol>
 
 
 
  <img src="https://static.igem.org/mediawiki/2015/4/4d/Ucl2015entrepprodthree.jpeg">
 
 
 
<h3> Why Chocolate? </h3>
 
<p>Independent studies have shown chocolate to be 3 to 4 times more effective in delivering probiotics to your
 
gut than dairy products, as it protects them from being degraded in the passage through the stomach.</p>
 
<p>It is a delicious treat that will not only benefit your gut flora, but also has all these added benefits:</p>
 
<ul>
 
  <li> <h4> Reduces the risk of cardiovascular disease </h4> </li>
 
  <p>There are hundreds of studies that provide supporting evidence that chocolate improves the functioning of the
 
    cardiovascular system, due to its high content in antioxidants.  </p>
 
  <p> The flavanols in dark chocolate have been shown to increase the production of Nitric Oxide (NO), which makes
 
    the arteries relax, lowering blood pressure, and facilitating blood flow (Fraga, 2011). This also improves
 
    athletic performance, and recovery due to its anti-inflammatory properties.</p>
 
<p>In addition to this, chocolate promotes the creation of good cholesterol, which in the long term reduces
 
      the cholesterol accumulation in the arteries. (Buijsse, 2006)</p>
 
  <li> <h4>Improve Brain function and enhance mood </h4> </li>
 
  <p> Dark chocolate contains compounds such as anandamide, PEA, theobromine and tryptophan, that increase serotonin
 
    and endorphin levels, which, in combination with our added probiotics, boost overall mood, and make you feel
 
    happier by encouraging the release of certain neurotransmitters such as melatonin. The consequences are increased
 
    libido, and an antidepressant effect.</p>
 
<p>The high flavonols content also improves memory and cognitive function, allowing you to be more aware, keep
 
      a sharp mind and a positive attitude (Scholey, 2013) </p>
 
  <li> <h4> It contains a high concentration of minerals </h4>
 
    <p>100g of Quality Dark Chocolate (70-85%) contains:</p>
 
<ul> - 67% of the RDA for Iron.</ul>
 
<ul> - 58% of the RDA for Magnesium. </ul>
 
<ul> - 89% of the RDA for Copper. </ul>
 
<ul> - 98% of the RDA for Manganese. </ul>
 
<ul> - plenty of potassium, phosphorus, zinc and selenium.</ul>
 
    <p>These are essential in building muscles, optimizing energy production, and are necessary for your
 
      immune function.</p>
 
</li>
 
</ul>
 
 
<h3> References </h3>
 
  <ul>
 
    <li>Buijsse, Brian, Edith JM Feskens, Frans J Kok, and Daan Kromhout. 2006. Cocoa intake, blood pressure, and
 
      cardiovascular mortality: the Zutphen Elderly Study. Archives of internal medicine 166, no. 4: 411-417.</li>
 
    <li>Fraga, César G, María C Litterio, Paula D Prince, et al. 2011. Cocoa flavanols: effects on vascular nitric
 
      oxide and blood pressure. Journal of clinical biochemistry and nutrition 48, no. 1: 63</li>
 
    <li>Scholey, Andrew, and Lauren Owen. 2013. Effects of chocolate on cognitive function and mood: a systematic
 
      review.Nutrition reviews 71, no. 10: 665-681.</li>
 
  </ul>
 
 
</div>
 
        </div>
 
 
 
        <div class="about-item3 hide">
 
          <div id="IP" class="protcl"><h2>Intellectual property</h2>
 
            <h4>
 
<p>From talking with our Business advisors, we realised that one of the main concerns that investors will have about
 
  us is the protection of our intellectual property, especially since the technology of our third phase product is
 
  going to be disclosed at our presentation. </p> </h4>
 
            <h3>Patent</h3>
 
<p>We are producing the first product of its type. While there are other probiotic companies that use a chocolate
 
  matrix as their delivery mechanism, they have only patented their own method, or their encapsulation process. We
 
  only found another similar patent, by P&G that patented the probiotics + chocolate combination, but it did as a
 
  method of promoting gastrointestinal health.
 
  <a href="http://worldwide.espacenet.com/publicationDetails/biblio?DB=EPODOC&II=2&ND=3&adjacent=true&locale=en_EP&FT=D&date=20120202&CC=AU&NR=2010264448A1&KC=A1%22%3E(Application%20number:%20AU20100264448%2020100624)">(Number: AU20100264448 20100624)</a> As a result,
 
  our invention is novel and non-obvious and we believe fully-patentable. </p>
 
<p>This gave us the opportunity to patent our method, in a similar way P&G did, but with the slight difference
 
  that its aim is mood and mental wellbeing enhancement.</p>
 
<p>The relevant documents can be found below</p>
 
<ul>
 
<li><a href="https://static.igem.org/mediawiki/2015/f/f0/Ucl2015entreppatentform.pdf ">Filled_Form.pdf</a></li>
 
<li><a href="https://static.igem.org/mediawiki/2015/0/0c/Ucl2015entreppatentreceipt.pdf ">Receipt.pdf</a></li>
 
</ul>
 
<p>Once we have developed the product further, to stages 2 and 3, we will be in the position to patent the specific
 
  engineered strains that we will add to upgrade our probiotic and make it serve the goal for which it was designed.</p>
 
  <h3> Trademark </h3>
 
  <p>During one of our policy and practice events, we were lucky enough to talk to some people with previous experience
 
  in the sector, and shared our prototype. They advised us to register our trademark as soon as possible, since we
 
  were distributing prototypes with our brand.</p>
 
<ul>
 
<li><a href="https://static.igem.org/mediawiki/2015/a/a3/Ucl2015entreptrade.pdf">Summary_of_trademark.pdf</a></li>
 
</ul>
 
 
<p>We expect the amount of IP to grow as we diversify and develop new products, and have appointed a specific
 
  department in our functional structure to take charge of it. This department will be specifically relevant
 
  in the third phase of development</p>
 
 
 
 
</div>
 
        </div>
 
 
 
<div class="about-item4 hide">
 
 
<div id="operations" class="protcl"><h2>Manufacturing Operations</h2>
 
    <div id="submenu" style="padding-top: 30px;">
 
        <a data-scroll data-options='{ "easing": "linear" }' href="#1" class="buttonblack">Prototype</a>       
 
        <a data-scroll data-options='{ "easing": "linear" }' href="#2" class="buttonblack">Production</a>
 
        <a data-scroll data-options='{ "easing": "linear" }' href="#3" class="buttonblack">Stratified</a>
 
      <a data-scroll data-options='{ "easing": "linear" }' href="#4" class="buttonblack">Personalised</a>
 
      <a data-scroll data-options='{ "easing": "linear" }' href="#5" class="buttonblack">References</a>
 
      <!--  <a data-scroll data-options='{ "easing": "linear" }' href="#4" class="buttonblack">Developers</a -->
 
    </div>
 
 
  <h3 id="1" style="padding-top:50px;">Prototype</h3>
 
  <p>With our product implementation plan in place, and a solid business model, we have developed a prototype
 
    with our engineered bacteria to test how effective the chocolate matrix is in encapsulating the bacteria and ensuring
 
  their survival for an effective delivery.</p>
 
 
 
  <h4>Protocol</h4>
 
<ol>
 
<li>Innoculate in two different Falcon tubes Nissle- TPH1(PSB1C3) 20ml of LB + Cam and shake overnight at 250rpm and
 
          37ºC</li>
 
<li>Centrifuge at 4000 rpm for 7min and discard the media.</li>
 
<li>Resuspend the pellet of one of the tubes in 1ml of a solution composed of 10mg of sugar and 1ml of miliQ water (A),
 
          and the one of the other in 1ml of miliQ water (B)</li>
 
<li> Melt 7g of dark chocolate (52%). The product also contained: Cocoa butter, Whey powder, Soya Lecithin,
 
        Polyglycerol polyricinoleate, and vanilla extract.</li>
 
<li>Mix 1g of chocolate with 500ul of the solution A (labeled 1.50 in the photo), and another 1g with solution B (labeled
 
          2.50). </li>
 
<li>Mix 1g of chocolate with 250ul of the solution A (labeled 1.25), and another 1g with solution B (labeled 2.25)</li>
 
<li>Mix 1g of chocolate with 500ul of mili Q water for the Control</li>
 
<p>Note: the chocolate must be at under 40ºC before mixing with bacterial solution</p>
 
<li>Leave samples in the fridge overnight</li>
 
      <figure>
 
      <center><img src="https://static.igem.org/mediawiki/2015/0/0a/Ucl2015entrepprototypesolid.jpg" style="width:564px;height:345px"></center>
 
      </figure>
 
        <li>Cut 0.5g of sample and wrap in aluminum foil for storage, to test shelf life</li>
 
      <figure>
 
      <center><img src= "https://static.igem.org/mediawiki/2015/5/5c/Ucl2015entrepprototypeshelfs.jpg" style="width:500px;height:381px"></center>
 
      </figure>
 
        <li>Cut 0.2g of sample and add to 5ml of LB in a falcon tube</li>
 
<li>Water bathe falcon tubes at 39ºC until the chocolate melts and mix by vortexing until a homogeneous
 
          texture is achieved</li>
 
<li>plate 100ul of solution in Cam LB agar plates and incubate overnight at 37ºC</li>
 
</ol>
 
<p> The bacteria grew back on the agar plates after having been placed on the chocolate matrix overnight, which
 
  showed chocolate is an adequate media to place them. In addition, we observed a higher number of CFU in the plate
 
  of solution B, suggesting the more effective method for resuspension of the pellet. </p>
 
  <figure>
 
  <center><img src= "https://static.igem.org/mediawiki/2015/4/4e/Ucl2015entrepprototypepetri.jpg" style="width:600px;height:373px"></center>
 
  </figure>
 
   
 
<p> However, from literature we know that the delivery and shelf life is best if the bacteria are microencapsulated
 
  beforehand with methods such as spray drying.  There is also literature showing that when tested in a simulator of
 
  the human digestive system, probiotics in a chocolate matrix proved to reach the gut more effectively than those
 
  in Dairy (Possemiers, 2010)</p>
 
<p> This is the first step towards a more sophisticated prototype, in which we will test freeze dried bacteria
 
  against other types of microencapsulation, to optimize the delivery, as we want to be able to guarantee that at
 
  least 2 billion CFUs reach the gut and colonize it. </p>
 
<p> Our finalized chocolate bar will be a 50g single dose rectangular bar. </p>
 
<figure>
 
  <center><img src= "https://static.igem.org/mediawiki/2015/f/f6/Ucl2015entrepprototypewrap.jpg" style="width:400px;height:444px"></center>
 
</figure>
 
     
 
  <h3 id="2">Scale-up Production</h3>
 
  <p>Manufacturing will be facilitated by outsourcing the production of our ingredients to key partners with
 
    experience in the sector. These partners will provide the already processed chocolate, and encapsulated
 
    probiotics. They will be selected according to the quality criteria with which the product identifies.
 
    This is mainly organic fair trade cocoa pods, and other more specific certifications to cater to the needs of
 
    specific sectors of the general public, such as a gluten free certification or dairy free.</p>
 
  <p>This will be beneficial to us, as these companies can off-set early production, which will allow our products
 
    to reach the market sooner. This allows us to focus on the further research of the biological components needed
 
    for the development of the company to the second and third stages.</p>
 
  <p> For the first stages of the business development, we would use strains whose effect is documented in literature</p>
 
  <figure>
 
  <center><img src= " https://static.igem.org/mediawiki/2015/d/d7/Ucl2015entreptablestrains.jpg " style="width:500px;height:446px"></center>
 
  </figure>
 
     
 
  <p>Production will take place following two different models, for two slightly different products</p>
 
  <h4 id="3"> Stratified </h4>
 
  <p>In the stratified model, consumers identify themselves with a specific category, and select the chocolate bar
 
    that is more adequate to suit their needs. </p>
 
<p> Whereas most probiotics companies we found offered a range of different products based on flavours we would
 
      target market segments instead. We will produce a fixed number of bars with specific probiotic contents,
 
      specifically designed for each market segment.  These will be differentiated in terms of age, sex, and
 
      lifestyle; which will lead to strata like: higher protein content for more athletic people,  or a limited
 
      amount of lactobacillus strains for vegans.</p>
 
<p>This model has a simpler production, given that it’s a repetitive process in which machines can be programmed
 
      for a certain amount of hours to produce one specific type of bar, and then switch to a different one by just
 
      changing the proportions to be added, and producing a great amount.</p>
 
 
 
  <h4 id="4"> Personalised </h4>
 
  <p>The personalized model, adds a layer of complexity to the structure. It will consist of made to order
 
      individual batches of probiotic chocolate tailored to the users goals and needs to in a way that has never
 
      been done before. </p>
 
<figure>
 
<img src="https://static.igem.org/mediawiki/2015/f/f9/Ucl2015entrepmicroprofile.jpeg" width="304" height="228">
 
        <figcaption>Simulation of customer's microbiome profile</figcaption>
 
</figure>
 
 
 
  <p> Using information supplied to us by the user we could design a probiotic fit specifically for their profile
 
      and monitor its effect by monitoring the state of the microbiome periodically. Over time more metrics and
 
      applications would be added, optimizing our ability to cater for each specific profile, and ultimately
 
      fulfilling the promise of power over all the potential that microscopic factories living within us can offer.</p>
 
 
 
  <h4 id="5"> References </h4>
 
  <ul>
 
      <li>Bravo JA, Forsythe P, Chew MV et al. (2011) Ingestion of Lactobacillus strain regulates emotional behavior
 
        and central GABA receptor expression in a mouse via the vagus nerve. Proc Natl Acad Sci USA 108, 16050–16055.</li>
 
<li>Dapoigny M, Piche T, Ducrotte, Lunaud B, Cardot JM, Bernalier-Donadille A (2012): Efficacy and safety
 
          profile of LCR35 complete freeze –dried culture in irritable bowel syndrome: A randomized , double blind
 
          study. World J Gastroenterol 18:2067-2075.</li>
 
<li>Desbonnet L, Garrett L, Clarke G et al. (2008) The probiotic Bifidobacteria infantis: an assessment of
 
          potential antidepressant properties in the rat. J Psychiatr Res 43, 164–174.</li>
 
<li>Hsiao EY, McBride SW, Hsien S et al. (2013) Microbiota behavioral and physiological abnormalities
 
          associated with neurodevelopmental disorders. Cell 155, 1451–1463.</li>
 
<li>Ko CY, Lin HTV & Tsai GJ (2013) Gamma-aminobutyric acid production in black soybean milk by Lactobacillus
 
          brevis FPA 3709 and the antidepressant effect of the fermented product on a forced swimming rat model.
 
          Process Biochem 48, 559–568. </li>
 
<li>Messaoudi M, Lalonde R, Violle N, Javelot H, Desor D, Nejidi A, et al. (2011): Assessment of
 
          psychotropic-like properties of a probiotic formulation (Lactobacillus helveticus R0052 and
 
          Bifidobacterium longum R0175) in rats and human subjects Br J Nutr 105:755-764 </li>
 
<li>Possemiers, Sam et al. (2010). Bacteria and chocolate: a successful combination for probiotic
 
          delivery."International journal of food microbiology 141.1: 97-103. </li>
 
<li>Rao AV, Bested AC, Beaulne TM, Katzman MA, Iorio C, Berardi JM, Logan AC (2009): A randomized,
 
          double-blind, placebo-controlled pilot study of a probiotic in emotional symptoms of chronic fatigue
 
          syndrome. Gut Pathog 1:6. </li>
 
      </ul>
 
</div>
 
</div>
 
 
<div class="about-item5 hide">
 
<div id="environment" class="protcl"><h2>Business Environment</h2>
 
  <div id="submenu" style="padding-top: 30px;">
 
        <a data-scroll data-options='{ "easing": "linear" }' href="#1" class="buttonblack">STEEPLE Analysis</a>       
 
        <a data-scroll data-options='{ "easing": "linear" }' href="#2" class="buttonblack">Regulations</a>
 
       
 
      <!--  <a data-scroll data-options='{ "easing": "linear" }' href="#4" class="buttonblack">Developers</a -->
 
    </div>
 
 
  <h3 id="1" ; style="padding-top:30px;">STEEPLE Analysis</h3>
 
<p>The STEEPLE framework is used to assess external factors in the business environment. In its original form it was conceived as the PEST analysis and accounted for political, economic, social and technological factors. In this analysis the augmented format is used, which includes legal, environmental and ethical factors. These are tightly interconnected and will often overlap.</p>
 
<p>In the following analysis we have not only outlined what external factors are likely to affect our business but also whether we expect to be positively or negatively impacted by it, much like in a SWOT (strengths, opportunities, weaknesses, threats) analysis.</p>
 
<br>
 
<h4>Social</h4>
 
 
<p>As mentioned previously, the improved perception of genetic engineering through the democratisation of the products and the industry itself will change people’s perceptions and thereby contribute to a better public image. These effects will in turn affect legislation and policy.</p>
 
<p>Stress is a constant of the human condition and, despite increases in material abundance, it has come to negatively impact health and quality of life more and more over the past decades. A full discussion of the reasons behind this phenomenon would yield a complex analysis unto itself. With more and more people suffering from chronic stress it has become imperative to find solutions for this problem that allow people to continue pursuing the lifestyles of their choice. </p>
 
<p>Health consciousness is a rising trend among peoples of all ages. This means people are more willing to spend more of their disposable income on health and wellness-related products. This has led to a surge in the probiotics market from () to () over () years. Given the familiarity of the public with probiotic products we can take them to a next level far more easily than if the concept were not already present in the public consciousness. The ageing population in the developed countries could present an opportunity in terms of targeting the elderly specifically. Given the fact that most older people take some form of medication our product would present a fresh alternative to another addition to a daily regimen of pills.</p>
 
<p>Attitudes toward mental health are slowly shifting towards having them considered to be on par with physical disorders in terms of their debilitating potential and the attention they deserve. The stigma that accompanies mental health is no longer as powerful and therefore people are more likely to seek assistance, including pharmacological interventions which we could supplement and, in the future, substitute.</p>
 
<p>The power balance between the doctor and the patient in the west is shifting in the direction of the patient and with this greater level of empowerment the patient is more likely to opt for a lifestyle product such as our minimum viable product (MVP) which can be given early on rather than a pharmacological treatment. </p>
 
<p>Safety and transparency have been prime concerns from day one. In order to gain and maintain the public’s trust we must not allow our company’s image to become associated with the idea of the impenetrable corporation with a dubious agenda that persistently clings to genetic engineering. </p>
 
<br>
 
<h4>Technological</h4>
 
 
<p>We are living in an unparalleled time in terms of scientific research and technological change. The cost of DNA sequencing is decreasing at a rate faster than Moore’s Law, and synthesis is also becoming much cheaper over time. The greater efficiency of new techniques allow us to gather information that was prohibitively expensive in the not-too-distant past, such as microbiome profiles and information about your personal genome. This information would be useless without the ability to use immense computer power and data science techniques to unlock powerful insights that would otherwise be unattainable. Recently developed techniques such as CRISPR/Cas9 will permit more accurate genetic manipulation of bacterial genomes in vivo, which will be crucial in developing our product strategy past the MVP.</p>
 
<p>Our MVP is based on a large and growing body of research in a transdisciplinary field that encompasses subjects ranging from genetics, to microbiology, to computer science to psychiatry. This allows us commission highly strategic studies to produce proprietary strains and patents which have information contained in the public domain as their foundation.</p>
 
<p>As the general public becomes more receptive to novel gadjets and the notion that technology can improve their lives we are witnessing a trend called the “quantified self”, characterised by devices that monitor several biometrics within the context of physical activity. This is not limited to performance athletes but also encompasses those who are both tech-savvy and focused on self-improvement. Since people in this group are likely to have large disposable incomes they could become the ideal target for our personalised MVP range.</p>
 
<br>
 
<h4>Economic </h4>
 
 
<p>The preponderant economic factor that influences the markets we’re looking at is the prospect of very low economic growth in the west that just doesn’t seem to be going away any time soon. To this we add the recent slowing down of emerging market economies.</p>
 
<p>Stagnant wages and low economic growth when coupled with rising inequality have led to a steady decrease in the disposable incomes of a the vast majority of the population and even a depletion of the middle class. This means it will be more difficult to draw these consumers into a long-term relationship with our product while it remains a wellness and lifestyle good; if it were seen as a medical treatment the price elasticity of demand  (in terms of % of income) would not be so large.</p>
 
<p>In response to the economic troubles of past years several countries have implemented policies of austerity which directly impact national health budgets. This reduces the market’s overall size but forces purchasers to become more creative in order to maximise their resources, thereby making them more open open to novel treatments with long-run cost-cutting potential. The search for creative solutions sparked by harsh economic times might come as a blessing in disguise to innovative endeavours such as this one </p>
 
<p>These conditions exacerbate the ever-present problem of a new technology offering potential benefits mainly to the most affluent. Not only is this undesirable in terms of the extent of the societal impact that we wish to make as a company but it also creates invokes unwarranted resentment towards technologies that should in principle become more affordable with time.</p>
 
<p>In spite, or perhaps because, of the current economic state of affairs the investment in biotech startups has only increased over the past few years. This trend is true of startups in general, but more pronounced within biotech. While some investors like Peter Thiel believe that it is with the startup innovation model that you can unlock the most value there are others who believe startups have become a bubble.</p>
 
<br>
 
<h4>Environmental</h4>
 
 
<p>The reasoning behind much of the tight regulation of products in this industry is the possibility that microorganisms will be released into the wild and the consequences of that happening. In the case of bacteria there is the possibility that genetically engineered plasmids will be taken up by wild bacteria which can then develop undesirable traits.</p>
 
<p>Given the poor environmental track record of genetically engineered seeds and other plant product we need to take steps to minimise our environmental impact not only for ethical reasons but also because of the public relations backlash that a poor environmental record entails.</p>
 
<p>The food ingredients should also be considered. Each element in our supply chain should be produce their contributions to our product in a sustainable and environmentally friendly way.</p>
 
<br>
 
  <h4>Political </h4>
 
 
<p>The shifting political landscape is likely to affect the genetic engineering aspects of our product. The public discourse on this issue has long been driven by interest groups on both sides and associated to large corporations sacrificing the public good for personal gain. Much of this is likely to change over the coming decade as patents on some of the original methods and products expire and the use of these technologies becomes democratized. As smaller companies run by young entrepreneurs enter the market it is likely that the image of genetic engineering will change to something more positive and relatable. Once these changes take place it will be possible for legislators beholden to their electorate to approach the problem of regulating this technology from a less biased standpoint. </p>
 
<p>Given the lack of public awareness of the general public with regard to this topic, educating the public about the benefits that synthetic biology could unlock should become one of our company’s priorities. Broadening this education to other areas in science would be even better.</p>
 
<p>The top-down hierarchy of the healthcare industry in general poses a significant obstacle to those looking to launch new products within the space. Since people do not possess the knowledge required to make informed medical decisions on their treatment they generally defer to a doctor on these matters. Because the doctor has several dozen (if not hundred) patients his willingness to take risks or try new products is far lower than if it were a patient ready to assume a risk for himself. It can be said that the incentives of doctors are not perfectly aligned with those of patients. This is not to say that doctors do not have their patient’s best interests at heart, but they also have to consider their own reputation and exposure to litigation. This effect becomes more pronounced as you move up the hierarchy of public health, thereby slowing down technical innovation tremendously.</p>
 
 
<br>
 
<h4>Legal</h4>
 
 
<p>As was discussed before the regulatory and legal constraints limit the range of products we can hope to put on the market at any point in time. We need to be aware of this from the start and do our utmost to comply with the safety and standards required by law at all times.</p>
 
<p>Different geographic regions pose different challenges when it comes to the legal aspect of the enterprise. The EU and the US tend to have the tightest regulatory structures, but since these are also the most desirable markets and those with which we have the greatest experience it would not serve our interest to use another body of legislation as a guideline. Therefore we have opted to use the highest standard we are aware of in order to be able to freely commercialize our product across all geographies with minimal effort.</p>
 
<p>This is not to say that a more progressive and less strict set of regulations in a particular country cannot provide us with the opportunity to prove that a particularly innovative product is safe and effective to use. This might be the case if we decide to create genetically engineered probiotics for non-medical use meant for the general public.</p>
 
<p>A source of opportunities that cannot be neglected is the possibility of particular molecular biology techniques being, because of their particular nature, permitted under the law. These methods would grant us greater flexibility in what we can do within the realm of consumer products in the near future.</p>
 
<p>Countries impose different restrictions on what can and cannot be patented within the domain of the life sciences. This is something we need to pay close attention to as we expand to other markets. The most important example of this is the 2013 US supreme court ruling that naturally occurring DNA sequences could not be eligible for patenting, thereby limiting what can be accomplished using only natural strains.</p>
 
<p>Given that we will be producing a food product we need to be aware of the limits to which we are liable for damages purportedly caused by the consumption of our products and what sort of safeguards we can install to minimize our legal risk. This isn’t restricted to the bacterial components in our product, but applies to all ingredients that could potentially be harmful to some people (allergens) as well as product contamination issues.</p>
 
 
  <br>
 
<h4>Ethical </h4>
 
 
<p>In analysing the business environment of a product such as this one the ethical aspect is something that cannot be overlooked. Several aspects have been been previously mentioned, including releasing patents on life, genetically engineered organisms into the wild, making safety and transparency front and center in the culture of our company, targeting people with mental health disorders and ensuring the supply chain is environmentally responsible.</p>
 
<p>The human rights aspects of the project need to be carefully considered. The supply chain should not only be environmentally responsible, but also socially responsible. Helping people and improving their quality of life are at the center of this enterprise, it’s ultimate goal is to provide those who have mental health an equal opportunity at a dignified life and the pursuit of their goals.</p>
 
<p>The mentally ill are one of the most disenfranchised groups in modern society, much more so because they have insufficient influential spokespeople for their cause. By helping the mentally ill we are attempting to correct one of the most jarring drivers of inequality in society: the arbitrary division of the sane and the insane.</p>
 
<br>
 
  <h3 id="2"> Regulations </h3>
 
 
 
<p> Prokao has been created under a complex structure, which means that each stream of development will be subject
 
  to different regulations. In general, it will be classified as a food, potentially entering the category that in
 
  the EU is regulated as  “novel foods”, as regulated by <a href="http://eur-lex.europa.eu/legal-content/EN/TXT/HTML/?uri=CELEX:31997R0258&from=EN">Regulation (EC) No 258/97 of the European Parliament and of the Council of 27 January 1997. </a>
 
<p>In addition to this, our third stage product might be classified as a GMO, in which case, the decision making
 
  process as to its commercialization and regulation, will be as follows:
 
<figure>
 
  <img src="https://static.igem.org/mediawiki/2015/7/74/Ucl2015entrepregulationsgmo00.jpeg">
 
  <figcaption>Source: <a href="http://ec.europa.eu/food/safety/index_en.htm">European Comission</a></figcaption>
 
  </figure>
 
 
 
 
</div></div>
 
 
<div class="about-item6 hide">
 
<div id="marketing" class="protcl"><h2>Marketing</h2>
 
<div id="submenu" style="padding-top: 30px;">
 
        <a data-scroll data-options='{ "easing": "linear" }' href="#1" class="buttonblack">Stratified - B2B</a>       
 
        <a data-scroll data-options='{ "easing": "linear" }' href="#2" class="buttonblack">Personalised - B2C</a>
 
      <!--  <a data-scroll data-options='{ "easing": "linear" }' href="#3" class="buttonblack">Developers</a -->
 
    </div>
 
<p> </p>
 
<h4 style="padding-top:50px;">Two Markets, Two Marketing strategies</h4>
 
 
<p>“The global functional foods market was worth an estimated USD43.27bn
 
in 2013. This figure has increased in value terms by 26.7% compared with
 
2009, and continues to demonstrate annual growth in excess of the world
 
food industry as a whole.” Market Report - Future Directions for the Global
 
Functional Foods Market - Leatherhead Food Research</p>
 
 
<p>Our minimum viable product(MVP) is a combination of probiotic strains and prebiotic substances that have
 
  been shown to promote mental wellbeing. Our vision is to target two very different but complementary markets
 
  by using two different approaches to sell our product. </p>
 
 
<h3 id="1">Stratified</h3>
 
<p>The first is the stratified approach, which consists of creating a range of probiotic chocolates designed to
 
  target particular groups of people. Differentiation on the base of age, gender, diet and body type means that we
 
  can create a product that is specific to the user while at the same time still being able to mass produce it. This
 
  business to business (B2B) wholesale strategy targets chains of shops that focus on a health conscious clientele
 
  such as Whole Foods in the US and Planet Organic in the UK as well as large retail stores with sections that cater
 
  to similar markets.</p>
 
 
<h3 id="2">Personalised</h3>
 
<p>The second, and most interesting, is the personalized approach. This product is the best way for the customer
 
  to extract the most benefit from our product, as it is tailored to him personally. Using a proprietary algorithm
 
  we would use information the patient provides about himself along with data from advanced diagnostic testing to
 
  design the best probiotic that current scientific knowledge allows us to create. The probiotic chocolate bars are
 
  made to order every month, with the information and recipe being updated accordingly. This product requires a
 
  certain degree of commitment because of the information that needs to be collected for the patient to take the
 
  most advantage of the product, notably regular microbiome sampling and analysis to provide information of the
 
  identity and size of different microbial populations in the gut.</p>
 
 
<p>Our marketing strategy is composed of two elements for each of our commercial approaches, the first being common
 
  to both and the second specific to the stratified or personalized approach.</p>
 
<p>The common strategy’s primary focus is brand awareness and the introduction of the concept of prebiotics as an
 
  aid to achieving and maintaining mental wellbeing. This involves participating in competitions, being featured in
 
  the media and engaging with social media constantly. Rather than intending to promote sales directly these
 
  endeavours are meant to increase the visibility of our product and company.</p>
 
 
<p>Our stratified range is sold to other businesses, so in order to increase our sales they need to be targeted as
 
  much as the final consumer. We will do this by attending events and fairs within the health and wellness industry
 
  and produce media content aimed at these businesses. We commit to active personal engagement and the formation of
 
  long term business partnerships with our most valued customers.</p>
 
 
 
<p>In order to increase the demand for our product we will target consumers while iterating through a variety of
 
  strategies, performing experiments designed based on lean management principles. Using targeted advertising and
 
  ads that feature different selling points we will be able to elicit the values of each market segment and find
 
  what is important to them (backed by scientific studies, ethical, fairtrade, green, etc.). We will also bring the
 
  product into public awareness by combining product placement with targeting influencers and thought leaders.</p>
 
<p>One of our strategies is to make consumers aware that Prokao is a healthy chocolate treat, something which we
 
  believe will be seen as highly desirable among both parents and their children. </p>
 
 
<p>Our personalised product targets a very narrow market of high value customers: those who are both willing to
 
  make permanent commitments in the name of their well being and those who can afford to do so. In an initial phase
 
  we will target customers that belong to specific interests groups by using podcasts and liaising with community
 
  leaders in order for word of mouth to produce viral growth. Features on specialty magazines and presentations at
 
  relevant conferences are also ways in which we can target this niche market. </p>
 
</div></div>
 
 
 
 
<div class="about-item7 hide">
 
<div id="management" class="protcl"><h2>Management</h2>
 
 
<h4>
 
<p>When it comes to small companies and startups, the organizational structure has to be designed to be the most
 
  efficient way for the company to achieve its goals and objectives. </p>
 
<p>Since our company has a long term plan of several parallel projects, we decided for a matrix structure where all
 
  can be worked on in a parallel way, without getting the streams of communication to interfere with each other. </p>
 
  <img src="https://static.igem.org/mediawiki/2015/c/c1/UCL2015entrepmanagement.jpeg">
 
  <h3>Functional Management</h3>
 
<p>We decided to give our management a functional structure, centered in job functions and departments, in blue.
 
    These will be overseen by the C’s suite. They will all report to the board of directors thus centralizing the
 
    decision making, which is essential in startups.</p>
 
<p>Functional structures are specifically designed to work in heavily project- focused companies, like ours.
 
  Directors will assign each of the projects to a manager, who will coordinate the whole development, getting
 
  support from each department in each aspect of the project development. This allows each project manager to be
 
  specialized in their stream and effectively meet deadlines, thus ensuring a more rapid business development,
 
  essential in the ever-changing conditions of the current market.</p>
 
 
 
  <h3>Matrix Organisation</h3>
 
<p>We specifically selected the matrix model because we consider that it’s the one that best allows the horizontal
 
    flow of skills and information, which, is essential in such a multidisciplinary endeavor. </p>
 
<p>It is common in the development phases, as it allows for large projects to move forward in a more
 
      organized way, since it allows employees to specialize in a project and apply their expertise in a particular
 
      field, without being removed from their department in question.</p>
 
<p>The project manager’s authority flows horizontally across departmental boundaries, but is always subjected
 
      to the board of directors.</p>
 
 
  <h3>Leadership</h3>
 
<p>In terms of leadership we have advocated for a Collaborative Leadership model, which encourages equal
 
      participation across all levels, therefore giving the project managers the most responsibility on the decision
 
      making and problem solving, almost at a CEO level.</p>
 
<p>This would imply open information sharing across the company, as every department as to be on the same
 
      page about each project, thus ensuring optimum efficiency in interdepartmental collaborations. This cross
 
      departmental participation allows more creative approaches to problem solving, and idea generation, speeding
 
      up once more business development.</p>
 
<p> Resource allocation will be managed by the board of directors. Team leaders will request these proactively
 
      and provide them to their teams, for which a fluent communication is key. This ensures a fast project advance
 
      as the employees have all resources available to do their jobs efficiently.</p>
 
<p>This type of structure places more responsibility on each team member, which makes them more involved in the
 
      process, and means issues are dealt with swiftly. The fact that leaders and team members are valued equally
 
      creates a better working environment and a better flow of feedback, which allows the team to be in a constant
 
      state of improvement; and employees to constantly acquire new knowledge and experience. This is essential to
 
      respond to the challenging and highly competitive environment that current market poses. </p>
 
<p>This structure responds to employees’ seek for autonomy and engagement, and promotes professional
 
      satisfaction, and a better environment for innovation. </p>
 
 
 
 
 
</ol>
 
</div></div>
 
 
<div class="about-item8 hide">
 
<div id="financialdata" class="protcl"><h2>Financial Data</h2>
 
<div id="submenu" style="padding-top: 30px;">
 
        <a data-scroll data-options='{ "easing": "linear" }' href="#1" class="buttonblack">Prototype</a>       
 
        <a data-scroll data-options='{ "easing": "linear" }' href="#2" class="buttonblack">Production</a>
 
      <!--  <a data-scroll data-options='{ "easing": "linear" }' href="#3" class="buttonblack">Developers</a -->
 
    </div>
 
 
  <p id="1">text</p>
 
<ol>
 
 
</ol>
 
 
</div></div>
 
 
<!--<div class="about-item9 hide">
 
<div id="iptg" class="protcl"><h2>IPTG-induced protein expression</h2>
 
<h4>
 
<ol>
 
 
<li>On the afternoon before the induction, start seed culture with appropriate antibiotic from glycerol stock and leave to incubate overnight at 37C shaking</li>
 
<li>The next morning, use 2 µl of seed culture to inoculate 100 ml of fresh media with appropriate antibiotic in shaker flask and grow until an OD600 nm of 0.4-0.6 is reached</li>
 
<li>If necessary, prepare these in the meantime :
 
<ul>- 50 ml stock of  lysis buffer (25 mM TRIS-Cl, 2 mM EDTA, pH 7.6):
 
<li> weight 0.151g of Tris base</li>
 
<li>add 45 ml of water</li>
 
<li>titrate with HCL to pH 7.6</li>
 
<li>fill up to 50 ml with water</li>
 
<li>add 29.2 mg of EDTA</li></ul>
 
<ul>- IPTG 100 mM stocks: (23.8 mg IPTG per 1ml ddH2O )</ul>
 
<li>When culture reaches OD of 0.4-0.6, add IPTG to a final concentration of 1 mM (=100ul of 100 mM stock)</li>
 
<li>Incubate induced culture at 30 °C for 4 hours</li>
 
<li>Split the culture into 4 falcon tubes (~25 ml each) and harvest the pellets by centrifugation for 20mins at max speed</li>
 
<li>Resuspend each pellet in 2 ml of  lysis buffer, lyse by sonication (10 cycles of 10 sec with 10 sec breaks)</li>
 
<li>Centrifuge 20 min at max speed.</li>
 
<li>Transfer all supernatants to separate tube</li>
 
<li>Measure the concentration of protein in supernatant using Bradford Assay</li>
 
<li>Store in the freezer</li>
 
 
 
 
 
</ol></h4>
 
</div></div>
 
 
 
<div class="about-item10 hide">
 
<div id="assays" class="protcl"><h2>Spectrophotometric assays</h2>
 
<h4>
 
<ol>
 
- Assay for tryptophan hydroxylase (<b>BBa_K1598002</b>) activity: <a href="https://static.igem.org/mediawiki/2015/c/cb/UCL_BBa_K1598002_Assay.pdf">download protocol</a>
 
 
</ol>
 
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Latest revision as of 01:59, 19 September 2015