Solution-phase binding of monoclonal antibodies to bee venom phospholipase A2.
The binding of monoclonal anti-bee venom phospholipase A2 antibodies to their antigen was monitored by size-exclusion high performance liquid chromatography. As judged by this panel of six antibodies, honeybee venom phospholipase A2 contains five binding sites, three of which are completely independent epitopes. The study revealed that this PLA2 can accommodate three different antibodies simultaneously. The results demonstrate the utility of size-exclusion high performance liquid chromatography in epitope analyses, such as its ability to compare the relative expansiveness and conformational state of the epitopes and to enumerate the antibodies that the antigen can accommodate simultaneously. The data provide compelling evidence that one of the monoclonal antibodies, M5 (which activates the enzyme), recognizes a different conformation of phospholipase A2 than do the other antibodies. The results also demonstrate that different pairs of monoclonal antibodies differ in their predilection to form high molecular weight complexes with the antigen.
Myatt
E A
EA
Center for Mechanistic Biology and Biotechnology, Argonne National Laboratory, IL 60439-4833.
Stevens
F J
FJ
Benjamin
C
C
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000378:metabolism*; D000937:Antigen-Antibody Reactions; D001514:Bee Venoms / Q000201:enzymology*; D001516:Bees / Q000201:enzymology; D046911:Macromolecular Substances; D051379:Mice; D008805:Mice, Inbred A; D010741:Phospholipases A / Q000276:immunology*; D054467:Phospholipases A2; D011485:Protein Binding; D012996:Solutions
D000911:Antibodies, Monoclonal; D001514:Bee Venoms; D046911:Macromolecular Substances; D012996:Solutions; D010741:Phospholipases A; D054467:Phospholipases A2