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Caltech chemists say antibody surrogates are just a 'click' away
Date:7/9/2009

des. "I actually traveled to Chicago with a vial of my capture agents as airline carry-on luggage, and came back with it, and the reagent still worked," says Agnew.

The technique makes use of the "in situ click chemistry" method, introduced by Sharpless in 2001, in which chemicals are created by joiningor "clicking"smaller subunits together.

To create a capture agent for a particular protein, the scientists devised a stepwise approach in which the first subunit of the capture agent is identified, and that unit, plus the protein, is used to identify the second subunit, and so on. For the first subunit, a fluorescent label is added to the protein, which is then incubated with a bead-based library of tens of millions of short-chain peptides, representing all the potential building blocks for the capture agent. When one of those peptides binds to the protein of interest, the fluorescent label is visualized on the bead (red, blue, or green, depending on the type of label), allowing the linked proteinpeptide complex to be identified.

That first peptidewhich is about a third of the length of the final capture agent the scientists are trying to makeis then isolated, purified, and modified on one end by the addition of a chemical group called an alkyne. This is the anchor peptide, which is then incubated, together with the same protein, with the bead-based library. The bead-based library now contains peptides that have been chemically modified to contain an azide group at one end. The alkyne group on the added peptide can potentially chemically react with the azide group of the library's peptides, to create a new peptide that is now two segments long.

However, the reaction can only occur when the second peptide comes into close contact with the first on the surface of the target protein, which means that both must have affinity for that protein; essentially, the protein itself builds an appropriate capture agent. The two-segment-long
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Contact: Kathy Svitil
ksvitil@caltech.edu
626-395-8022
California Institute of Technology
Source:Eurekalert

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