title: Protein Assembly in Serum and the Differences from Assembly in Buffer.
abstract: This chapter illustrates how analytical ultracentrifugation methods, coupled with the fluorescence detection system, are an excellent approach to characterizing and comparing protein-binding interactions in dilute solution and concentrated, crowded solutions like serum. We show that in serum, the binding and assembly states for a pair of endogenous protein ligands and an antibody inhibitor are dramatically different than those observed in dilute, simple buffers. This type of analysis approach may be helpful in research efforts intent at discerning the underpinnings to a therapeutic's activity and pharmacokinetic properties in vivo.
authors.affiliation: Amgen, Seattle, Washington, USA. Electronic address: hilljb@u.washington.edu.
authors.forename: John J
authors.initials: JJ
authors.lastname: Hill
authors.affiliation: Center to Advance Molecular Interaction Science, University of New Hampshire, Durham, New Hampshire, USA.
authors.forename: Thomas M
authors.initials: TM
authors.lastname: Laue
mesh_terms: D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000302:isolation & purification; D002021:Buffers; D006801:Humans; D053244:Osteoprotegerin / Q000737:chemistry / Q000302:isolation & purification; D011485:Protein Binding; D055503:Protein Multimerization; D053245:RANK Ligand / Q000737:chemistry / Q000302:isolation & purification*; D044967:Serum / Q000737:chemistry; D012709:Serum Albumin / Q000737:chemistry / Q000302:isolation & purification; D012996:Solutions; D014461:Ultracentrifugation / Q000379:methods
chemical_list: D000911:Antibodies, Monoclonal; D002021:Buffers; D053244:Osteoprotegerin; D053245:RANK Ligand; D012709:Serum Albumin; D012996:Solutions; C505688:TNFSF11 protein, human
keywords: Analytical ultracentrifugation; Antibody; Assembly; Binding; Clusters; Concentrated solution; Confined; Crowded; Fluorescence detection system; Osteoprotegerin; RANKL; Sedimentation velocity; Serum