Separation of monoclonal antibodies from antihapten antisera by two-dimensional affinity electrophoresis.
A high-resolution two-dimensional affinity electrophoresis (2D-AEP) method was developed, using capillary polyacrylamide gel (PAG) isoelectric focusing in the first and slab PAG affinity electrophoresis in the second direction. Using this method, anti-hapten antibodies were separated into a number of monoclonal antibody [immunoglobulin G (IgG)] families, each of which is composed of several IgG spots having an identical affinity to the hapten but different isoelectric points. 2D-AEP may offer a powerful tool for solving fundamental problems in immunochemistry such as antibody heterogeneity, its hapten binding specificity and antigen-dependent somatic mutation.
Takeo
K
K
First Department of Biochemistry, Yamaguchi University School of Medicine, Ube, Japan.
Nakamura
K
K
Suzuno
R
R
D000911:Antibodies, Monoclonal / Q000302:isolation & purification*; D003429:Cross Reactions; D003619:Dansyl Compounds / Q000737:chemistry; D004140:Dinitrophenols / Q000276:immunology; D015180:Electrophoresis, Gel, Two-Dimensional; D006241:Haptens / Q000276:immunology*; D007525:Isoelectric Focusing; D012116:Resins, Plant
D000911:Antibodies, Monoclonal; D003619:Dansyl Compounds; D004140:Dinitrophenols; D006241:Haptens; D012116:Resins, Plant