Dye-hydrophobic hapten conjugate/anti-dye antibody complex as immunogen: preparation of hydrophobic hapten-specific monoclonal antibodies.
In order to induce the production of antibodies specific for small molecules, it is common to link them to a protein. However, when the small molecule is very hydrophobic it is extremely difficult to prepare such a conjugate. Here, we describe a simple way to obtain an antigenic conjugate under controlled conditions: in a first step a very hydrophobic hapten, cholanic acid, is linked to a dye, basilen blue, in organic solvent; in a second step the cholanic acid-basilen blue conjugate is dissolved in phosphate buffered saline and mixed with rabbit polyclonal anti-basilen blue antibodies previously raised in rabbits against basilen blue-key-hole limpet hemocyanin conjugate. Such a complex, which dissociates very slowly, appears to be a good immunogen in mice. Anti-cholanyl residue monoclonal antibodies were produced and characterized.
Chevrie
K
K
Centre de Biophysique Moléculaire, CNRS, Orléans, France.
Motta
G
G
Mayer
R
R
Monsigny
M
M
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry*; D000918:Antibody Specificity; D000936:Antigen-Antibody Complex / Q000737:chemistry / Q000276:immunology*; D002793:Cholic Acids / Q000737:chemistry; D006241:Haptens / Q000276:immunology*; D051379:Mice; D011817:Rabbits; D014227:Triazines / Q000737:chemistry
D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D002793:Cholic Acids; D006241:Haptens; D014227:Triazines; C012842:Cibacron Blue F 3GA; C007036:cholanic acid