am working on SPR, i would like to know the permanent fixation or immobilization of antibody on the CM5 gold chip. by usual immobilization wizard the antibody is easily removed after regeneration. thanks inadvance.
I would suggest that you modify the protocol available at http://www.ncbi.nlm.nih.gov/pubmed/18274533
In this paper the authors describe how to take glycosylated antibodies, convert the sugar into a dithiol group, and then use the dithiol group to mediate a strong attachment to a gold surface (in their case, nanoparticles). The modification chemistry is very simple, but be aware that the Sensopath linker is not soluble in water - it should be resuspended in something like ethanol, DMF, or DMSO. I have used ethanol and DMF with good success.
Hi there, see my publication for more details where I used EDAC/NHS successfully to immobilize antibodies against whey proteins.
Hi, you can self assemble alkyl thiols with carboxylic acid as functional group on gold surface and after that by using EDC/NHS, immobilize your antibody.
Self-Assembled Monolayers (SAM) is a great way to attach proteins or ABs to gold surfaces (search on google scholar: "sam on gold"). RM Corn has done a lot in spr field. Google scholar search: "spr corn"
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