Report
Teaser, summary, work performed and final results
Periodic Reporting for period 1 - IMPAM (Development of a new immunoassay platform based on porous antibody microparticles)
Teaser
The efficiency and at the same time price availability of in vitro tests systems for analyte determination are one of the main critical needs in medical and environment diagnostics, for example for the control of toxins, cancer markers, markers of inflammation, etc. The modern...
Summary
The efficiency and at the same time price availability of in vitro tests systems for analyte determination are one of the main critical needs in medical and environment diagnostics, for example for the control of toxins, cancer markers, markers of inflammation, etc. The modern research directs on reaching these tasks. The traditional and widespread approach in the field of diagnostics (enzyme-linked immunosorbent assay, ELISA) has sensitivity limitations due to the use of planar supports for the detection of analytes. Besides that, the reduction of the amount of expensive reagents, such as antibodies, required to perform reliable testes, is crucial to make these tests readily available. In this project, we developed a new approach for biosensing based on porous antibody-contained microparticles that addresses these issues.
The comparison of the new version of an immunoassay designed by us with traditional ELISA showed the following advantages of our system: 1) a higher sensitivity and 2) reducing the amounts of antibodies required to perform assays. Thus, the crucial tasks were solved. We believe that our new approach for the determination of analytes can expand the range of users, being useful in a wide diapason of analytical applications, and will be implemented in medical practice.
Thus, the aims declared for the action were achieved.
Work performed
The following results were achieved during the action:
1) Different methods of the creation of antibody containing protein particles were established and their usefulness was compared. All methods were based on the CaCO3 particle fabrication. The method of co-precipitation was chosen as the most perspective and effective one, and then optimized. It was shown that antibody containing protein particles keep their structure after CaCO3 - template dissolving and, thus, can be used for the further immunoassay designs.
2) The new format of immunoassay, based on antibody containing protein particles, was developed for the determination of high and low molecular weight analytes. For this aim we used human Immunoglobulin G as a marker of the infections and Ochratoxin A as a fungal toxin, respectively.
3) The advantages of the new assays were demonstrated in comparison with traditional ELISA. The sensitivity of the immunoassay for the determination of human Immunoglobulin G was increased by a factor of 10. At the same time, the amount of needed antibodies was reduced by the same factor. The comparison of our format of immunoassay with the traditional ELISA for the determination of Ochratoxin A showed increasing of the sensitivity and decreasing of used concentration of antibodies by the factor of 3.
For the dissemination of results they were presented at two conferences, Biosensors 2016 (Gothenburg, Sweden) with a poster presentation and 3rd BiPoCo2016 (Szeged, Hungary) with the oral one. Thus, the certain number of scientists and potential users of diagnostic tests became acquainted with our research.
Moreover, during the action a training visit to Nottingham Trent University (UK) for exchanging experience and knowledge took place. The possibility of the fabrication of CaCO3 crystals with adjusted porosity through soft templating was estimated during this period.
The results received during the action are being prepared for publications . Drafts of the publications are attached to Part B.
Final results
Our project combined knowledge from different disciplines. For the first time the idea of creation of protein microparticles (technique from material science) was applied towards immunology/biotechnology. This allowed developing a new approach of immunoassay for solution of diagnostic tasks in medical and environmental areas. The advantages of the approached immunoassay systems were contained in the increasing of sensitivity and reduction of the reagent’s amount. Thus, this could improve the potency of diagnostic tests at the same time increasing their economical availability. These both parameters within the framework of modern life will bring benefits to the sector of healthcare. If to express it in figures, researchers will spend from 3 to 10 times (depending on the type of antibodies) less expenses on reagents. As it was said above, it leads to a cheaper diagnostic analysis combined with its increased sensitivity. As a result, using these diagnostics tests will be available for a wider number of people.