Project overview: The DiaCHIP

The DiaCHIP is an innovative tool to simultaneously and differentially detect antibodies present in blood sera in response to an infectious disease. It bears the potential to greatly simplify broad band screenings, detection of autoimmune diseases and the determination of vaccination statuses. Especially in case of threatening infections accompanied by similar symptoms fast and reliable differentiation could save lives.
To account for a disease by verification of corresponding antibodies in a patient’s blood serum is a well-known and routinely used system in modern diagnostics (LINK – diagnostics today). Thus, a simple blood sample is sufficient to perform reliable diagnostics with the DiaCHIP.
The key feature in the concept of the DiaCHIP is the combination of on-demand protein synthesis and a novel, label-free detection method in one device. This enables to overcome challenges in storage and handling that occur with currently available tests. Additionally, diagnoses can be received faster and in a more cost-efficient way than until now. Nonetheless, the whole device is so simple that it can easily be rebuilt and utilized by future iGEM teams (LINK – new device).

Step 1: Preparing the DiaCHIP by protein synthesis

Before being able to screen for antibody-antigen interactions, antigens have to be synthesized and immobilized in a microarray arrangement. This is obtained by a copying mechanism transforming a DNA template into a protein microarray by cell-free protein expression. This expression system based on a bacterial lysate prevents the need for genetically engineered organisms to produce every single antigen.
The template slide is placed in close proximity to the future protein array enabling the expressed proteins to reach this other surface by diffusion. Complex chemistry ensures that target proteins are specifically immobilized on this surface, while components of the expression mix can be washed away before sample analysis. To obtain the DNA template, the respective sequences containing transcriptional and translational initiation sites, the antigen coding sequence and terminating regions have to be constructed and labeled with an amino group. An activated PDMS slide provides the basis for immobilization of the DNA by covalent binding of the amino group. Spotting the antigen coding sequences in a distinct pattern enables to retrace a detected binding event to a certain disease.

Step 2: Measuring serum samples using iRIf

After preparation of the DiaCHIP, a patient’s serum sample can be flushed over the protein array. The binding of antibodies to the protein surface causes a minimal change in the thickness of the slide right at the corresponding antigen spot. This change can be measured without the need for a further label with a new method called iRIf (imaging reflectometric interference). Based on the interference of light beams reflected on different thin layers, binding events can be recorded in real-time.

After weeks of opimizing the different components of the DiaCHIP, we reveal our great results. The highlight of our project was reached with the successful detection of antibodies in our own blood!