title: An indirect method to measure trimerization constants using surface plasmon resonance.
abstract: The ability of surface plasmon resonance to precisely measure kinetic binding constants was exploited here to indirectly evaluate the thermodynamic dissociation trimerization constant (K(d)) of a designed chimeric protein, IZN-23, derived from an isoleucine zipper and a portion of the N-terminal helix residues of HIV-1 gp41. The method uses two monoclonal antibodies (mAbs) that display different off-rates when binding the monomeric or trimeric IZN-23. A detailed description of the data analysis strategy employed to unravel the K(d) trimerization constant from the observed off-rate kinetic values is presented, and the potential exploitation of this technique in different fields is highlighted.
authors.affiliation: Biotechnology, IRBM, 00040 Pomezia, Italy.
authors.forename: M
authors.initials: M
authors.lastname: Mattu
authors.forename: C
authors.initials: C
authors.lastname: Bendtsen
authors.forename: G
authors.initials: G
authors.lastname: Barbato
mesh_terms: D015700:HIV Envelope Protein gp41 / Q000032:analysis* / Q000378:metabolism; D015497:HIV-1 / Q000378:metabolism; D016350:Leucine Zippers; D055503:Protein Multimerization; D011994:Recombinant Proteins / Q000032:analysis / Q000378:metabolism; D020349:Surface Plasmon Resonance / Q000379:methods*
chemical_list: D015700:HIV Envelope Protein gp41; D011994:Recombinant Proteins