We are looking for an easy and effective method to adsorb/immobilize proteins on a PMMA surface. We already tried oxygen plasma treatment, however, the efficiency of the coating is not sufficient for our application. Thanks for your help in advance...
1. Change plasma parameters. It will work. Even air corona discharge. Nitrogen in the gas will improve binding strength.
2. PMMA is easily hydrolyzed. Proceed with the surface cooh groups according to the books.
Dear Gerhard,
Thanks for your help.... Actually, we have improved now the plasma parameters (by increasing power and time, according to Tsougeni et al.,
Microelectronic Engineering, 2009) and the first results look very promising. However, the idea with adding Nitrogen into the plasma process sounds very interesting and we will try this as well.
We also thought about the option using directly a EDC/SNHS chemistry after hydrolization (as suggested in Point 2). But, we would like to have an easy protein coupling method with less chemistry (e.g. only adsorption) because of our final application.
Best regards, Can
Just an update: meanwhile we optimized the required plasma parameters (power, time and gas composition) for surface activation of different polymers, and also compared it with EDC/SNHS activation. Using oxygen plasma treatment, the antibody adsorption works very well, please see attached picture for the results on PMMA. Details will be published hopefully soon...
Update: We have published now the results of our plasma activation study on polymeric surfaces (especially, PMMA foils) in Polymers (MDPI): https://www.mdpi.com/2073-4360/12/1/104/pdf
Dear Can Dincer,
That's great work!
Congratulations!
I think a further study is required to elucidate the orientation aspect of antibodies especially in the formation of sandwich structures both on solid phase and solution phase. In addition, its significance may further clear via the reduction of Ab consumption in related immunoassays besides its significant role in simplifying the conjugation process.
Thanks for your kind comment. Glad that you liked our work!
Regarding the orientation of antibodies, you're right that it could be further studied. However, in our current work, we focus on a sensor application using a competitive assay where Au-NP-labelled antibodies only employed for optical signal generation. Hope to present this study within this year as well...
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