Team:Michigan/Project/

From 2014.igem.org

(Difference between revisions)
 
(14 intermediate revisions not shown)
Line 2: Line 2:
<html>
<html>
<body>
<body>
 +
<div style="background-color:white;position:absolute;height:25px;width:250px;top:5px;left:0px"></div>
<div id = "main-wrap">
<div id = "main-wrap">
Line 25: Line 26:
<div id = "header-container" class = "container">
<div id = "header-container" class = "container">
-
<div id = "logo">
+
<div id = "logo" style="width:150px;height:150px">
 +
 
<a id = "site-title" href = "index.html">
<a id = "site-title" href = "index.html">
-
<img src = "http://che.engin.umich.edu/enewsletter/winter2013/images/MSBTlogo2.jpg" alt = "MSBT" />
+
<img src = "https://static.igem.org/mediawiki/2014/6/61/Umichsigil.jpg" alt = "MSBT" style="width:110px;height:110px;position:absolute;top:-76px;left:1px" />
</a><!--- #site-title -->
</a><!--- #site-title -->
<span id = "site-description">
<span id = "site-description">
-
2014 Michigan Synthetic Biology Team
 
</span><!--- #site-description -->
</span><!--- #site-description -->
</div><!--- #logo -->
</div><!--- #logo -->
Line 45: Line 46:
<ul class = "sub-menu">
<ul class = "sub-menu">
<li class = "nav-menu-item">
<li class = "nav-menu-item">
-
<a href = "elements.html">Background</a>
+
<a href = "https://2014.igem.org/Team:Michigan/Project/">Background</a>
</li>
</li>
<li class = "nav-menu-item">
<li class = "nav-menu-item">
-
<a href = "columns.html">Design</a>
+
<a href = "https://2014.igem.org/Team:Michigan/Design/">Design</a>
</li>
</li>
<li class = "nav-menu-item">
<li class = "nav-menu-item">
-
<a href = "columns.html">Wet Lab</a>
+
<a href = "https://2014.igem.org/Team:Michigan/Modeling/">Modeling</a>
</li>
</li>
<li class = "nav-menu-item">
<li class = "nav-menu-item">
-
<a href = "columns.html">Dry Lab</a>
+
<a href = "https://2014.igem.org/Team:Michigan/Results/">Results</a>
</li>
</li>
<li class = "nav-menu-item">
<li class = "nav-menu-item">
-
<a href = "columns.html">Future Plans</a>
+
<a href = "https://2014.igem.org/Team:Michigan/Future/">Future Plans</a>
</li>
</li>
<li class = "nav-menu-item">
<li class = "nav-menu-item">
-
<a href = "columns.html">Accomplishments</a>
+
<a href = "https://2014.igem.org/Team:Michigan/Accomplishments/">Accomplishments</a>
</li>
</li>
Line 73: Line 74:
<li class = "nav-menu-item">
<li class = "nav-menu-item">
-
<a href = "portfolio_filterable.html">Human Practice</a>
+
<a href = "https://2014.igem.org/Team:Michigan/Human/">Human Practice</a>
<ul class = "sub-menu">
<ul class = "sub-menu">
<li class = "nav-menu-item">
<li class = "nav-menu-item">
-
<a href = "#">Outreach</a>
+
<a href = "https://2014.igem.org/Team:Michigan/Outreach/">Outreach</a>
</li>
</li>
<li class = "nav-menu-item">
<li class = "nav-menu-item">
-
<a href = "#">Econmic Analysis</a>
+
<a href = "https://2014.igem.org/Team:Michigan/Economic/">Economic Analysis</a>
</li>
</li>
<li class = "nav-menu-item">
<li class = "nav-menu-item">
-
<a href = "portfolio_filterable.html">Covance Partnership</a>
+
<a href = "https://2014.igem.org/Team:Michigan/Covance/">Covance Partnership</a>
</li>
</li>
<li class = "nav-menu-item">
<li class = "nav-menu-item">
-
<a href = "portfolio_filterable.html">Collaborations</a>
+
<a href = "https://2014.igem.org/Team:Michigan/Collaborations/">Collaborations</a>
</li>
</li>
Line 97: Line 98:
<li class = "nav-menu-item">
<li class = "nav-menu-item">
-
<a href = "blog.html">Notebook</a>
+
<a href = "https://2014.igem.org/Team:Michigan/Notebook/">Notebook</a>
-
<ul class = "sub-menu">
+
-
<li class = "nav-menu-item">
+
-
<a href = "blog.html">Winter Planning</a>
+
-
</li>
+
-
+
-
<li class = "nav-menu-item">
+
-
<a href = "blog_single.html">Project Journal</a>
+
-
</li>
+
-
</ul>
+
</li>
</li>
Line 127: Line 119:
</div><!--- #header-container -->
</div><!--- #header-container -->
</header><!--- #header -->
</header><!--- #header -->
-
+
<!--- <div style="background-color:#f1ed6f;width:994px;height:120px;position:absolute;left:-5px;top:51px"></div> -->
Line 134: Line 126:
<header id = "page-header" class = "center">,
<header id = "page-header" class = "center">,
-
<h1><font size="10" <em class = "colored">About our Project: OsmY</em></font></h1></p><font size="3"><font color="#191970"> 2014 Michigan Synthetic Biology Team </font></font color></p>
+
<h1><font size="10"> Background for our Project</font></h1></p><font size="3"><font color="#191970"> University of Michigan iGEM Team </font></font color></p>
 +
 
</header><!--- #page-header -->
</header><!--- #page-header -->
Line 142: Line 135:
<!-- put information  because this is a link to this page -->
<!-- put information  because this is a link to this page -->
-
    <p>Many important pharmaceutical protein, such as antibodies and insulin, are derived from mammalian organisms. These mammalian cells have organelles that isolate a special environment for proteins to fold correctly and form disulfide bonds, such as the endoplasmic reticulum. Yet, mammalian cells take a long, difficult time to grow  and are not ideal for industrial production of pharmaceutical proteins. Fast growing organism such as <i>E.coli</i> presented us with the grand advantage of producing and purifying large amounts of protein in a short amount of time but prokaryotes don’t have the organelles capable of properly folding pharmaceutical proteins.
+
<div style="background-color:white;height:1px"></div>
-
    </p><br><p>
+
-
    Disulfide bonds occur naturally in <i>E.coli</i> in the periplasm. The periplasm has an oxidizing environment, similarly to the endoplasmic reticulum, that allows disulfide bond formation. If we drive a mammalian protein through the periplasm of an E.coli cell it will able to fold properly by disulfide bond formation.
+
-
</p><br><p>
+
-
    Antibodies have become paramount in biotechnology and medicine. Table 1 ranks the companies with the highest sales in March 2014; as we can see, 3 companies out of the top 10 sell antibody fragments. This reveals the impact of purification system improvement.
+
-
    </p><br><p>
+
-
    In our project, we've purified antibody fragments by tagging a secretion protein, OsmY, to an antibody scFv (single chain fragment variable) and over-expressing the construct in <i>E. coli</i>. To demonstrate the importance of our study we've chosen an antibody specific to DADH, a Salmonella protein. Salmonella is responsible for 23,000 hospitalizations in the US each year, highlighting the need for large quantities of inexpensive sensors. To further demonstrate the value of our part we've compared our secretion system with industrially used secretion system that use the secretion tag pelB. </p><p></p>
+
-
                                                </div>
+
-
<div style="height:200px"> <img src="https://static.igem.org/mediawiki/2014/8/8e/Background_image.png" style="height:275px;width:400px;position:absolute;top:455px;left:100px">  
+
-
<p> <font style="position:absolute;top:430px;left:250px"> Table 1 </font></p>
+
-
<p> <font size="1" style="position:absolute;top:660px;left:500px"> "List of Therapeutic Monoclonal Antibodies." Wikipedia. Wikimedia Foundation, 13 Oct. 2014. Web. 14 Oct. 2014.
+
-
"Salmonella." Centers for Disease Control and Prevention. Centers for Disease Control and Prevention, 28 Aug. 2014. Web. 14 Oct. 2014.</p>
+
<p>Many important pharmaceutical proteins, such as antibodies and insulin, are derived from mammalian organisms. These cells have an organelle called the endoplasmic reticulum that creates an isolated environment for proteins to fold correctly and form disulfide bonds. Yet, mammalian cells can be difficult to grow and are not ideal for industrial production of pharmaceutical proteins. Fast growing organisms such as E.coli provide us with the advantage of producing and purifying large amounts of protein in a short amount of time. However, prokaryotes don’t contain endoplasmic reticulums, making it challenging to properly fold pharmaceutical proteins.</p></br>
-
</div>
+
 +
<p>Disulfide bonds occur naturally in E.coli in the periplasm, the space between the inner and outer membranes of the bacteria. The oxidizing environment of the periplasm is similar to the endoplasmic reticulum and allows disulfide bond formation. Indeed, funneling a mammalian protein through the periplasm of an E.coli cell allows for proper folding and disulfide bonding.</p></br>
-
                            <!--- #page-content -->
+
<p>Additionally, the production of antibodies have become paramount in biotechnology and medicine. As you can see in Table 1, of the top 10 drugs with the highest grossing sales in 2014, three of the drugs are comprised of purified monoclonal antibody therapies. Antibody therapies remain difficult to produce and purify, and thus improvements on the purification process are greatly needed to continue to produce these drugs efficiently.  </p></br>
 +
 
 +
<p> In our project, we've created a construct capable of purifying antibody fragments by tagging a bacterial secretion protein, OsmY, to an antibody scFv (single chain fragment variable) and over-expressing the construct in E.coli. To demonstrate the importance of our study we've chosen to produce an antibody specific to Dihydroxy acid Dehydradatase (DADH), a Salmonella protein. Salmonella infections are responsible for 23,000 hospitalizations in the US each year, highlighting the need for large quantities of inexpensive sensors and treatments. To further demonstrate the value of our part we've compared our secretion system with industrially used secretion system that use the secretion tag pelB. </p>
 +
                                              <div> </div>
 +
<div style="height:200px"> <img src="https://static.igem.org/mediawiki/2014/8/8e/Background_image.png" style="height:275px;width:391px;position:absolute;top:480px;left:293px">
 +
<p> <font style="position:absolute;top:465px;left:472px"> Table 1 </font></p>
 +
 
 +
<p style="position:absolute;top:497px;width:100px;left:177px"> <font size="1"> "List of Therapeutic Monoclonal Antibodies." Wikipedia. Wikimedia Foundation, 13 Oct. 2014. Web. 14 Oct. 2014.</font></p></p>
 +
 
 +
 
 +
</div>
</div><!--- #page-container -->
</div><!--- #page-container -->

Latest revision as of 03:29, 18 October 2014

<! -- Michigan Synthetic Biology Team -->

This is an announcement, click the Close button on the right and it will not show again until you restart your browser.

Close

Many important pharmaceutical proteins, such as antibodies and insulin, are derived from mammalian organisms. These cells have an organelle called the endoplasmic reticulum that creates an isolated environment for proteins to fold correctly and form disulfide bonds. Yet, mammalian cells can be difficult to grow and are not ideal for industrial production of pharmaceutical proteins. Fast growing organisms such as E.coli provide us with the advantage of producing and purifying large amounts of protein in a short amount of time. However, prokaryotes don’t contain endoplasmic reticulums, making it challenging to properly fold pharmaceutical proteins.


Disulfide bonds occur naturally in E.coli in the periplasm, the space between the inner and outer membranes of the bacteria. The oxidizing environment of the periplasm is similar to the endoplasmic reticulum and allows disulfide bond formation. Indeed, funneling a mammalian protein through the periplasm of an E.coli cell allows for proper folding and disulfide bonding.


Additionally, the production of antibodies have become paramount in biotechnology and medicine. As you can see in Table 1, of the top 10 drugs with the highest grossing sales in 2014, three of the drugs are comprised of purified monoclonal antibody therapies. Antibody therapies remain difficult to produce and purify, and thus improvements on the purification process are greatly needed to continue to produce these drugs efficiently.


In our project, we've created a construct capable of purifying antibody fragments by tagging a bacterial secretion protein, OsmY, to an antibody scFv (single chain fragment variable) and over-expressing the construct in E.coli. To demonstrate the importance of our study we've chosen to produce an antibody specific to Dihydroxy acid Dehydradatase (DADH), a Salmonella protein. Salmonella infections are responsible for 23,000 hospitalizations in the US each year, highlighting the need for large quantities of inexpensive sensors and treatments. To further demonstrate the value of our part we've compared our secretion system with industrially used secretion system that use the secretion tag pelB.

Table 1

"List of Therapeutic Monoclonal Antibodies." Wikipedia. Wikimedia Foundation, 13 Oct. 2014. Web. 14 Oct. 2014.