A synthetic polypeptide conjugate from a 42-residue polypeptide and salicylhydroxamic acid binds human myeloperoxidase with high affinity.

摘要: This thesis describes the design and synthesis of small molecule derivatives their polypeptide conjugates as high affinity binders for proteins: D-dimer protein (D-dimer), a biomarker diagnosis thromboembolic diseases; human myeloperoxidase (MPO), cardiovascular chitinases, potential targets asthma therapy. The interactions between synthetic binder molecules those proteins were evaluated by surface plasmon resonance (SPR) biosensor analysis fluorescence spectroscopy. Competition SPR experiments or other methods proved that components critical binding specifically bound to original site molecules. consisted 42-residue helix-loop-helix conjugated via aliphatic spacers suitable length. could be any type moderately structure. In development D-dimer, tetrapeptide GPRP with dissociation constant Kd 25 μM was used 4C15L8GPRP obtained estimated approximately 3 nM. MPO, salicylhydroxamic acid (SHA) 2 4C37L34C11SHA 0.4 theobromine derivative (pentoxifylline) 43±10 4C37L34-P considerably higher than pentoxifylline. identified from 16-membered pool candidates conjugating each member set 16 designed polypeptides. affinities greatly enhanced 2-3 orders magnitude, compared molecule. polypeptides did not bind measurable affinities. discovery these new can pave way diagnostic tests in vivo vitro, independent antibodies.