well, I want to conjugate my 3 peptides(with mW: 150, 62, 28 KDa) separately to PLGA(50:50 with mW:30000-60000), and i will use carbodiimide method for this conjugation. But in this process, the question is: how can i find how much peptide and how much PLGA can i react together to have high output?? which ratio is good?? Is there any suitable and easily solution?
Hi Ruhollah Vasegh, you have to optimize your conjugation concentration, you can start with 2:1 w/w of PLGA to peptides ratios..carbodiimide activation reaction is pH dependent so be keep your MES (Morpholino ethanesulphonic acid) buffer pH between 5.4 to 6.2, second thing is PLGA, take acid terminated PLGA instead of ester terminated PLGA as carbodiimide activates the COOH group present in the PLGA, if you are using PLGA nanoparticles for conjugation then its batter to use surfactant like pluronic f 68 for particle formation which has COOH group, which can enhance your conjugation to particle surface. use NHS as intermediate stabilizer during PLGA carbodiimide reaction...you can refer the following paper for conjugation
Nanoparticles Targeting Dendritic Cell Surface Molecules Effectively Block T cell Conjugation and Shift Response
Chuda Chittasupho,1 Laura Shannon,2 Teruna J. Siahaan,1 Charlotte M. Vines,2 and Cory Berkland1,3
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Dear Jairam Meena,
Thanks for your answer, but about ratio that you suggest, i think we should use peptide excess (more than PLGA) to have high output, not PLGA 2 time more than peptide, because if we do, at the end we have amount of PLGA that doesn't conjugate and make our out put too low... isn't it?
yes you are right Ruhollah Vasegh, your peptide should be in excess for high peptide to polymer ratio
dear follower, well, i think i find the solution...
check this article out, it shows how much PLGA and reagent we can react...
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