The whole world solemnized New Year with great enthusiasm and energy. This day has a special contingency for people. People concerned to various walks of life welcome New Year in their own hearty ways. It is a day which expresses joy and happiness all around. Genuine success comes only to those who are ready for it. So never step back and always have courage to accept new challenges. May all of you get succeed in the year 2018 and achieve all your goals you have set.
Since my last blog, I was doing bureaucracy work and literature review. The aim of my project is to “develop the platform for multiplexed detection of antibiotics in food sample based on DNA-addressable arrays with smartphone read out system“.
Herein, I would like to share the short description of my research work. I started the experimental work for the selection of most suitable antibiotics with high consideration of their relevance in real food sample. Firstly, I selected tylosin as a target antibiotic because it was first evaluated at the twelfth meeting of committee for veterinary medicinal product in 1968 when it was concluded that tylosin used in animal feed or in veterinary medicine should not give rise to detectable residue in edible products.
Tylosin is a macrolide antibiotic that is active mostly against Gram-positive bacteria and mycoplasmas. It is ineffective against Enterobacteriacea. It consists of predominantly tylosin A (factor A), but varying amount of tylosin factor B (desmycosin), tylosin factor C (macrosin), and tylosin factor D (relomycin) may also present, depending on the manufacturing source.
The three different antibodies were available for the detection of tylosin in our laboratory. Four different bioconjugates (BSA-conjugates) was prepared for the development of an immunoassay against these antibodies, hTA-BSA, hTB-BSA, TA-BSA, and TB-BSA.
All combinations were tested by ELISA. From the results, we came to conclusion that hTB-BSA is most suitable bioconjugate with its highest affinity against clone C126.96.36.199. The detectability achieved in terms of IC50 was in the nM range (ppb).
For future perspective, according to the DNA-direct immobilization protocol. A set of four complementary pairs of ssDNA (N4-N8, up and down) were being synthesized. These sequences have 50% G/C content (purines/pyrimidines) and have the same composition but in different sequences. Furthermore, we will employ this strategy for the detection of multiplex antibiotics with openSPR system to validate this study. Finally, we will use smartphone interface system to make the system more compact for hand on use.
OpenSPR is the benchtop instrument. It provides high quality, label-free interaction analysis for a fraction of the cost of existing solutions. OpensPR is user friendly and sets up in just minutes, so we can start generating data right away. This unique nano-structured sensor surface delivers repeatable, highly sensitive kinetic data for proteins, antibodies, nucleic acids, and more.
Moreover, during this period I got the chance to visit South Korea for some bureaucracy work where I had met with my friends and recall my old memories. The weather condition was worst for me during my stay as compared to Barcelona.
Recently, Jordi Neils, ESR3, has joined our group for his secondment. Hopefully, we will have a good time with him. I also will get the chance to work with him on LSPR-based analysis.
Hopefully, all of you will enjoy my blog. Good luck for your project.