Team:ZJU-China/Demonstrate
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OVERVIEW
Our work is to Create an artificially constructed multi-enzyme complex that constructs a logic gate at the protein level! We made the first generation of products, A detector I, and achieved very satisfactory results, improved our product and get A detector II, tested it in the hospital and implemented our design
(See more in our Project design-Logic Gate)
Based on protein structure prediction and sequence analysis, we DIY some enzymes ourselves. They are linked together by the Tag-Catcher system and form a multi-enzyme complex. Through the relationship between the substrate and the product, they form well-run logic gates. This is one of the highlights of our project - the protein logic circuit!
Fig. 1
Protein modeling constructed by de novo modeling Robetta.
Fig. 2
SDS-PAGE results indicated that the three enzymes were successfully expressed.
A detector Ⅰ
(See more in our Electrode)
By using the multi-enzyme complex immobilized on a electrode and an electrochemical workstation, we constructed an enzyme electrodes, which we called A detector Ⅰ！This sensor has a unique way to immobilize the enzymes - matrix formed by the Curli protein. This method is not only robust, but also capable of retaining enzyme activity. The result of the single enzyme test and the detection of logic gates are both met our expectation well.
Fig. 3
Quantitative Congo Red (CR) Binding Assays to verify the successful expression of Curli.
Fig. 4
The current-time curve shows that A detector 1 distinguishes different inputs well
A detector II
(See more in our Applied Design)
Although the results of the A detector Ⅰare satisfactory, through many expert consultations and field research(See more in our integrated Human Practice), We found that the product may encounter difficulties in practical use.. Therefore, we designed the second generation of products - A detector II! Compared to the huge size of the A detector I, the A detector II is so small that it can even be carried in your pocket! We used A detector II to test the patient's blood sample in the hospital and found that it can successfully distinguish the blood samples of patients and normal people, and give the results within 30s.
Fig. 5
Our own designed interdigital electrodes. Print the multi-enzyme complex onto the electrode by 2D printing.
Future work
Improve user experience with A detector II, and make UI more user-friendly
Use A larger sample size to train A detector II. The way of machine learning will make the test results more accurate
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