title: Antigen-binding site protection during radiolabeling leads to a higher immunoreactive fraction.
abstract: It is generally accepted that the immunointegrity of an antibody (Ab) depends on the preservation of its antigen-binding sites. Our goal was to radiolabel an antibody at several iodine:antibody molar ratios under conditions protecting its combining site and to compare its immunoreactive fraction (IRF) and electrophoretic mobility with those of the same antibody radiolabeled without protection. The data indicate that an antibody radiolabeled while its antigen-binding site is occupied by its antigen had the same IRF, regardless of the number of iodine atoms per antibody molecule. On the other hand, even at an I:Ab ratio of 1:1, the IRF of the same antibody radiolabeled without protection was lower than that of a protected one and decreased with increasing I:Ab ratios. In addition, the iodination of these Ab changes their electrophoretic mobility; however, when the Ab is labeled in the protected state, the degree of change is less. The binding of an antibody to its antigen prior to radiolabeling, therefore, enhances its immuno-integrity and prevents major conformational changes as reflected by electrophoresis.
authors.affiliation: Department of Radiology (Nuclear Medicine), Harvard Medical School, Boston, Massachusetts 02115.
authors.forename: A D
authors.initials: AD
authors.lastname: Van den Abbeele
authors.forename: R A
authors.initials: RA
authors.lastname: Aaronson
authors.forename: S
authors.initials: S
authors.lastname: Daher
authors.forename: R A
authors.initials: RA
authors.lastname: Taube
authors.forename: S J
authors.initials: SJ
authors.lastname: Adelstein
authors.forename: A I
authors.initials: AI
authors.lastname: Kassis
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000937:Antigen-Antibody Reactions / Q000276:immunology*; D001666:Binding Sites, Antibody / Q000276:immunology*; D004587:Electrophoresis, Agar Gel; D006041:Goats / Q000276:immunology; D007457:Iodine Radioisotopes; D007553:Isotope Labeling / Q000379:methods; D051379:Mice; D011817:Rabbits
chemical_list: D000911:Antibodies, Monoclonal; D007457:Iodine Radioisotopes