Using a technology based upon protein microarrays, a core screening technology, allowing thousands of samples to be printed or spotted onto an ordinary glass slide, was invented back in the nineteen eighties. This development in custom antibodies and research antibodies has provided groundbreaking advances in the field. A glass slide is treated with aminosilane or another protein binding surface chemistry, and then the slide is coated with the target protein necessary for the creation of custom antibodies. This is the protein chip.
The culture supernatants are then printed onto the protein chip and incubated for one hour to allow any custom anitbodies present to bind with the protein. The only research antibodies remaining on the surface of the protein chip are the research antibodies that specifically recognise the target protein. The slide is then incubated with isotype specific secondary anti-mouse custom antibodies. Each of the isotype specific anti-mouse secondary custom antibodies has a different flourophore. The secondary research antibodies bind to the target-specific custom antibodies on the protein chip.
The slide is then washed and scanned in a microarray scanner and different colours are visualised. The presence of a spot indicates the presence of target-specific research antibodies. The colour of the spot indicates the isotype of the research antibodies. Because each culture supernatant can be screened against tens or even hundreds of target antigen chips, it is possible to map where on the protein the custom antibodies bind. It can also discriminate between research antibodies that may recognize phosphorylated or non phosphorylated forms of the same target antigen.
It is, of course, essential to generate high-affinity custom antibodies using novel formulations and immunisation protocols which are both effective and humane. A good sign that custom antibodies (aka) research antibodies are of exceptional quality is whether or not the adjuvant components have been licensed for use in humans, which would suggest that the custom antibodies are consequently very mild and yet extremely effective.