to cross-link a protein to a polysaccharide, which to choose? Sulfo-NHS or NHS? - (Apr/24/2008 )
hi all,
ahh..just, may I ask that, when trying to conjugate a protein with a polysaccharide by using the zero-length cross-linking procedure, which chemical will most labs choose, NHS or sulfo-NHS, together to work with EDC? And why will it be chosen?
P.S. Is there any classical paper about carbohydrate-protein conjugate synthesis recommended?
thanks!
a stupid undergraduate
ahh..just, may I ask that, when trying to conjugate a protein with a polysaccharide by using the zero-length cross-linking procedure, which chemical will most labs choose, NHS or sulfo-NHS, together to work with EDC? And why will it be chosen?
P.S. Is there any classical paper about carbohydrate-protein conjugate synthesis recommended?
thanks!
a stupid undergraduate
I assume you will do the reaction in water/buffer. NHS or Sulfo-NHS does not make any difference. This method is only good for a polysaccharide that has groups like COOH.
ahh..just, may I ask that, when trying to conjugate a protein with a polysaccharide by using the zero-length cross-linking procedure, which chemical will most labs choose, NHS or sulfo-NHS, together to work with EDC? And why will it be chosen?
P.S. Is there any classical paper about carbohydrate-protein conjugate synthesis recommended?
thanks!
a stupid undergraduate
I assume you will do the reaction in water/buffer. NHS or Sulfo-NHS does not make any difference. This method is only good for a polysaccharide that has groups like COOH.
Yes, the reaction will be done in an aqueous buffer, and the carbonhydrate wight be either heparin or heparan sulfate. But I'm just a bit wandering that, if considering sulfo-NHS gets an -SH group that NHS doesn't have, will there be any differences in their water solubility that might influence their effeciency in the procedure? .........thanks a lot for replying the question
If you are doing real reaction, not for you course work, you can do a quick experiment as follows: dissolve some NHS and sulfo-NHS in buffer that you use. The main point for using sulfo-NHS is its improved solubility in water. NHS is very soluble in water already. Some jerk invented this problem to get a research paper, I guess. You end up paying much more $$ for that stupid chemical.
IT's true! The price of sulfo-NHS is much more expensive, and that' why my professer want me to make sure which to use before ordering. So, when only considering the solubility, NHS will be effective enough to carry out the conjugation reaction and the difference in solubility won't contribute to a significance difference in the experiment results, right?
Thanks a lot for providing such an important information! And just because I haven't buy the chemicals yet, or I'll try the experiment you mentioned. thanks again!
no difference in performance. Order from Sigma/aldrich, its is way cheaper.