title: Immunochemical quantification of procion red HE-3B used as ligand in affinity chromatography.
abstract: The quantification of Procion Red HE-3B used as a ligand in affinity chromatography for proteins is reported. It's based on an enzyme-linked immunosorbent assay using antibodies against the dye. Polyclonal antibodies were classically prepared after conjugation of the dye on KLH and injection into rabbits. The development of the assay was based on the competitive inhibition between hemoglobin-dye complex and free dye. The sensitivity of this method was about 1000-times higher than a classical spectrophotometric assay, and was modulated by some chemical substituents attached on the native dye. It was demonstrated that the assay was applicable to the determination of dye traces that may be released from dye affinity sorbents. Moreover, the quantification of the dye was successfully applied to proteins that are being purified from a dye affinity column.
authors.affiliation: IBF S.A., Sepracor Bioprocessing division, France.
authors.forename: P
authors.initials: P
authors.lastname: Santambien
authors.forename: P
authors.initials: P
authors.lastname: Girot
authors.forename: I
authors.initials: I
authors.lastname: Hulak
authors.forename: E
authors.initials: E
authors.lastname: Boschetti
mesh_terms: D002846:Chromatography, Affinity; D004396:Coloring Agents; D004797:Enzyme-Linked Immunosorbent Assay; D007120:Immunochemistry; D008024:Ligands; D015394:Molecular Structure; D010958:Plasminogen / Q000032:analysis; D011506:Proteins / Q000032:analysis*; D012680:Sensitivity and Specificity; D013053:Spectrophotometry; D014227:Triazines
chemical_list: D004396:Coloring Agents; D008024:Ligands; D011506:Proteins; D014227:Triazines; C020795:procion red HE-3B; D010958:Plasminogen