title: Screening of protein-ligand interactions by affinity chromatography.
abstract: This paper examines affinity chromatography (AC) as an alternative tool for the determination of protein-ligand interactions for the particular case in which the ligand is the same protein. The methodology is less labor-intensive and more sample-efficient than traditional methods used to measure the second virial coefficient (B(22)), a parameter commonly used to evaluate protein-protein interactions. The chromatographic capacity factor (k') was studied for lysozyme and equine serum albumin for a wide range of experimental solution conditions such as crystallizing agent concentration, protein concentration and pH. Parallel experiments using AC to determine k' and static light scattering (SLS) to determine B(22) showed that the two parameters were highly correlated. Two different column volumes ( approximately 1 and approximately 0.1 mL) were tested and gave essentially the same values for k', showing the feasibility of miniaturization.
authors.affiliation: Department of Chemistry, 118 Hand Chemical Laboratory, Mississippi State University, Mississippi State, Mississippi 39762, USA.
authors.forename: Carlos D
authors.initials: CD
authors.lastname: García
authors.forename: Steven C
authors.initials: SC
authors.lastname: Holman
authors.forename: Charles S
authors.initials: CS
authors.lastname: Henry
authors.forename: W William
authors.initials: WW
authors.lastname: Wilson
mesh_terms: D002846:Chromatography, Affinity / Q000295:instrumentation* / Q000379:methods*; D002919:Chymotrypsinogen / Q000737:chemistry*; D019281:Dimerization; D019544:Equipment Failure Analysis; D005240:Feasibility Studies; D006863:Hydrogen-Ion Concentration; D008024:Ligands; D046911:Macromolecular Substances; D008904:Miniaturization; D009113:Muramidase / Q000737:chemistry*; D011485:Protein Binding; D011506:Proteins / Q000737:chemistry*; D012996:Solutions
chemical_list: D008024:Ligands; D046911:Macromolecular Substances; D011506:Proteins; D012996:Solutions; D002919:Chymotrypsinogen; D009113:Muramidase