Production of antibodies against degradative neoepitopes in aggrecan.
The use of synthetic peptides to generate rabbit polyclonal anticatabolic neoepitope antibodies that can be used to study the presence of defined proteolytic cleavage sites in aggrecan is described. Principles of peptide design and methods for preparation and characterization of ovalbumin conjugates are presented along with approaches for the characterization and affinity purification of the resulting antisera. Limitations associated with the use of antipeptide antibodies to study authentic protein neoepitopes are discussed.
Mort
John S
JS
Joint Diseases Laboratory, Shriners Hospital for Children, and Departments of Surgery and Medicine, McGill University, Montreal, Quebec, Canada.
Roughley
Peter J
PJ
D053674:Aggrecans; D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000032:analysis / Q000276:immunology; D000939:Epitopes / Q000737:chemistry / Q000276:immunology*; D016326:Extracellular Matrix Proteins / Q000737:chemistry / Q000276:immunology*; D007106:Immune Sera / Q000096:biosynthesis* / Q000302:isolation & purification; D037181:Lectins, C-Type; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry / Q000276:immunology*; D011509:Proteoglycans / Q000737:chemistry / Q000276:immunology*; D011817:Rabbits; D013388:Succinimides / Q000737:chemistry
D053674:Aggrecans; D000906:Antibodies; D000939:Epitopes; D016326:Extracellular Matrix Proteins; D007106:Immune Sera; D037181:Lectins, C-Type; D010455:Peptides; D011509:Proteoglycans; D013388:Succinimides; C049271:N-succinimidyl bromoacetate