Hope so, all of you will be doing great with your research. As all of you know that in FoodsmartPhone project a stags comes several times, where we have to think innovatively to solve concerned issues in order to move forward in your research.
In this blog I would like to emphasize on “how we can be more innovative and creative in research”. More preciously, our thinking also needs to be well-informed, organized, matured, and even saturated within your professional context. But on its own, knowledge of a field without the stimulus of an effort to rebuild to vary an assumption, will always just produce more of the same.
It’s worst to miss one good idea for fear of writing down half-baked ones, a timidity problem that seems to acutely affect many intellectuals. As William Emmerson remarked: ‘In every work of genius we recognize our own rejected thoughts; they come back to us with a certain alienated majesty’.
Edward de Bono wrote, a whole (rather dreadful) book about the difficulties people have in thinking about what is only ‘possibly correct, possibly wrong’.
Scientific progress often involves an ability to notice small (often minutely) unusual outcomes. A first stage in seeing a new idea that we prosaically explain in unreflective ways, seeing past ‘common sense’ accounts of how things work. Thomas Edison said “keep on the lookout for novel ideas that others have used successfully”, your idea has to be original only in its adaptation to the problem you’re working on. Develop your feelings, and constructively use your professional commitments. The creative idea and push it a bit further drivers, so long as your powers of realistic critique are not too dulled. Use analogies, metaphors, images, little prototypes to drive intuitive explanation. By repeating in imagination, we de-sense what has been given to the senses.
Martin Heidigger argued: ‘The anxiety of those who are intellectually daring cannot be opposed to joy or even to the comfortable enjoyment of tranquilized bustle.
Next, I would like to tell you about my ongoing research, as I mentioned in my previous blogs the objective of my projected is to develop innovative platform to detect target antibiotics by employing DNA-directed immobilization (DDI) strategy with optical read out system which finds its potential application in real sample, which would be a milk in this case.
Amongst four bio-conjugates, which we synthesized (TA-BSA, TB-BSA, hTA-BSA, and hTB-BSA), hTA-BSA and hTB-BSA were most suitable bio-conjugates and have highest affinity against clone C220.127.116.11 out of three clones of TylB, C18.104.22.168, C22.214.171.124, and C126.96.36.199 for the detection of target antibiotic (TylB). We have developed competitive assay by using two target antibiotics, TylA and TylB against hTB-BSA and hTA-BSA with clone C188.8.131.52. On the experimental and theoretical study, we concluded that hTB-BSA/C184.108.40.206 is the most suitable combination for the detection of TylB by taking the advantage of its less haptem cost and important analytical parameters such as, IC50, maximum absorbance value, and regression coefficient value.
Furthermore, we analyzed the matrix effect to validate the potential of this strategy for TylB detection in fresh milk. PBST buffer absorbance value was chosen as a reference, the trend of graph confirmed that milk would have to be diluted to the reach the reference signal.
Furthermore, we have developed two dimensional ELISA assay to determine the optimized concentration of hTB-BSA against clone C220.127.116.11 for TylB detection in milk and validated it by competitive assay with two analytes, TylA and TylB in real sample. For the proof of concept, hTB-BSA/C23.4.24 vs TylA and TylB was tested in milk sample (Figure 2D) and we have achieved IC50 in nanomolar range (ppb) for selected combination of hTB-BSA/C6.23.4.
Next, we have designed the DDI strategy for the conjugation of hTB with specific oligonucleotide (hTB-N4up) for the capturing of target antibiotic. Following sequence has been employed in this study, [N4up:5’-(AmC6) CTCTGTACACTAACGCTGGA-3’]. NAP10 column (sephadex G-25, DNA grade) was used to isolate the hTB-N4up from reaction mixture and purified by HPLC. The difference in mass peak between control (N4up) and hTB-N4up confirmed that hTB was conjugated successfully with N4up. For the proof of concept, the collected sample was further analyzed by MALDI-TOF mass spectrometry which also confirmed the conjugation of hTB with N4up.
In future, we will employ this highly selective and selective strategy for the detection of target antibiotics with OpenSPR system, works on the principle of LSPR. hTB-BSA will be captured by the sensors ship which have pattern gold nanostructure on it. The flow of fluid contains hTB-BSA through PDMS microchannel binds on the sensor chip. After capturing hTB-BSA, we will employ clone C18.104.22.168 to confirm the binding phenomenon by online monitoring through customized Nicoya software. The regeneration will be done through sodium hydroxide- based buffer medium.
Moreover, I would like to say something about family, family dynamics are tricky, especially as you grow up and embark on your own life. But it’s important to appreciate your family’s quirks and treat your time together (and away from work!) as an opportunity to relax and renew, rather than seeing it as a source of stress. Yes, right now I am in my home town and can’t explain my feeling to see my family members.
Hope to see you all in summer school at Prague, Czech Republic.