antigen / antibody immobilization - (Feb/19/2003 )
My field of work is devices and we are working with molecule-device hybrid structures.. i have little knowledge in microbiology.. i would like to gain some understanding of protein binding to functionalized substrates:
1. best ways to bind antigen (or antibody ) or protein molecules to a substrate like glass with SiOx surface, to substrates that are parafinic in nature say surfaces attached with long chain alkanes etc. (i haven't found good literature on it).. or suggest any other substrate functionalization that would be ideal for such attachment ?
2. are antigen molecules and antibody molecules active even when attached as above ? ie after drying in N2 atmosphere etc.. does an antibody (suspended in solution) attack an antigen so attached (antigen is attached to surface and dried) ? or does the antigen need to be in solution ?
3. Selectivity of antibody attack, what is the mechanism of identifying ? my concern is do they attack inorgonic or organic substrates on which I attach antigen.. say substrates that are parafinic, epoxy based, glass like etc. how are the charge transfers during the reaction between antibody and antigen.. ?
comments ? experiences ? or references are welcome.. thanks in advance.
look like you are developing protein array. It's not easy to coat protein on glass surface, proteins are not like nucleic acids they are much more delicate, sensitive to conformational change and other micro-environmental conditions. It's more easy to coat enzyme substrates or antibodies on chip since they are more homogeneous in terms of structure. The trick is to let them to stand up on chip (using a common chemical treatment on glass) to be able to capture their targets. the rest is easy....Looking around how many HD (high-density) protein arrays/tissue arrays are available? you can get the idea.