Team:GreatBay United/Contribution

• Home
• Project
  1. Background
  2. Description
  3. Design
  4. Contribution
• Wetlab
  1. Proof of Concepts
  2. Engineering
  3. Result
  4. Measurements
  5. Parts
  6. Safety
  7. Lab Notebook
• Drylab
  1. Hardware
  2. Modeling
  3. Proposed Implementation
  4. Supporting Entrepreneurship
  5. Sustainability
• Human practices
  1. Integrated Human Practice
  2. Education
  3. Education and Communication
  4. Excellence in Other Area
• Team
  1. Team
  2. Attibution
  3. Collaboration
  4. Partnership

• Part usage by GBU
• Part:BBa_K416000
• 3D Modeling

Contribution

GreatBay_United would also like to pass the spirit of helping future iGEM teams on. So we completed component information and added the information to existing component pages. This year, we used yeast surface display technology in our project, and Part:BBa_K416000 was used as the signal peptide. Our relevant results have been updated on the page of Part:BBa_K416000. What's more, we also updated existing hardware and software, corrected errors in documents and made 3D printed hardware files. In the process of hardware design, we used 3D printing technology to complete our hardware. We summarized the experience related to 3D modeling in the process of design and implementation, and we hope to provide useful references for iGEM team in the future.

Part:BBa_K416000 Usage by GreatBay_United

In 2021, GreatBay_United has used this part to display their protein complex onto the yeast surface.
The protein expression cassette was showed below:

The expression cassette was successfully transformed into yeast, and the relevant proteins were successfully expressed, see the fluorescence image below:

In another cassette, GFP antibody (anti-GFP) were also expressed, if both complex and antiGFP were successfully displayed on the surface, they will be able to sense the HCV NS3 protease in solution. The protease would cut and release the antiGFP linked to GFP, and the solely expressed anti-GFP would had chance to connect with GFP. this interaction would lead to the macroscopical agglutination phenomenon when we mix two kinds of engineered yeast together and also HCV protease. The final agglutination could also prove that the signal peptide is working well.

The image below shows the two types of yeast co-culture with HCV protease in the medium

The image below shows the two types of yeast co-culture without HCV protease in the medium

3D Modeling

We design and produce a piece of hardware to better apply the use of artificial limulus amebocyte lysate to our daily life. By simply filling the inner tank with tap water and following the steps in instructions, patients will be able to detect endotoxin level of bacteria by themselves. In order to produce a prototype that can reveal our aim to the greatest extent, we make assumptions and do trials and errors. After looking into many different types of technique to produce the hardware, we decide to use 3D printing technique. The following contents are 3D modellings of the hardware and some of our experience.

The slotted edge of the inner tank makes sure that the lid can perfectly embedded the top of the inner tank. The slight bulge at on the lid helps people open it. During the 3D painting process, several generations of models are designed. The first one only consists of a small inner tank, and heating plate is designed to be outside the tank. However, since Abs plastic has poor heat conductivity, the model is abandoned. The second version contains a heating plate inside, but the resistance of the plate is too high that the adapter does not fit the plate. Also, the layout is not concise enough, and the inner tank will sometimes leak. For the final version, the heating plate is replaced by one with appropriate resistance, and we used glass glue instead of heat melt glue to avoid leaking. The 3D model is also redrawn to fit the new heating plate.

The slotted edge of the inner tank makes sure that the lid can perfectly embedded the top of the inner tank. The slight bulge at on the lid helps people open it. During the 3D painting process, several generations of models are designed. The first one only consists of a small inner tank, and heating plate is designed to be outside the tank. However, since Abs plastic has poor heat conductivity, the model is abandoned. The second version contains a heating plate inside, but the resistance of the plate is too high that the adapter does not fit the plate. Also, the layout is not concise enough, and the inner tank will sometimes leak. For the final version, the heating plate is replaced by one with appropriate resistance, and we used glass glue instead of heat melt glue to avoid leaking. The 3D model is also redrawn to fit the new heating plate.

We design and produce a piece of hardware to better apply the use of artificial limulus amebocyte lysate to our daily life. By simply filling the inner tank with tap water and following the steps in instructions, patients will be able to detect endotoxin level of bacteria by themselves. In order to produce a prototype that can reveal our aim to the greatest extent, we make assumptions and do trials and errors. After looking into many different types of technique to produce the hardware, we decide to use 3D printing technique. The following contents are 3D modellings of the hardware and some of our experience.

1. At first, we failed to print the bottom the inner box because we did not level the 3D printing machine. The second stage is the same as the first stage.

3. After leveling the machine, the box was printed, but we found the lid of the box was too hard to take off.

4. We designed a little bulge to make the box easier to be opened. At this stage, we successfully made the inner box, but considering the poor heat conductivity of Abs plastic, we then decided to put the heating plate inside the inner box.

5. We created two small slots for the heating plate, but this kind of structure was too difficult to be sealed. No matter how hard we tried, the slot would leak.

6. We used another structure to avoid leaking, but it did not work. Also, the resistance of the heating plate did not fit the voltage provided by the adapter, which will be discussed in the modeling part.

7. Now we started to work on the big box. This was the version of big box, containing a window to show the screen of temperature controlling chip. However, we did not take the volume of the chip into consideration, so the big box is not big enough.

8. The big box is now big enough, but we met some technical problems. The Abs plastic could be used to print such a big box. It would collapse. This box was actually made of many small pieces of plastic, and we used glue to stick them together. In addition, we could not increase the thickness of the box because it would take too much time for printing.

9. The same as 8, but it broke.

10. In the end, the inner box was printed successfully with the best structure for avoiding leaking. What is more, we used glass glue instead of heat melt glue as well to seal the box. The big box was also completed using PLA plastic, which was more suitable for bigger objects.