Generation of monoclonal antibodies specific for mouse apolipoprotein B-100 in apolipoprotein B-48-only mice.
Over the past 10 years, many laboratories have investigated lipid metabolism and atherogenesis with a variety of transgenic and gene knockout mouse models. Although many of these studies have yielded valuable insights, some have been hampered by a paucity of useful antibodies against mouse proteins. For example, many laboratories have analyzed genetic and dietary interventions affecting lipoprotein metabolism without useful antibodies against mouse apolipoprotein (apo) B. In this study, we sought to develop highly specific monoclonal antibodies against mouse apoB-100. To achieve this goal, gene-targeted mice that synthesize exclusively apoB-48 (apoB-48-only mice) were immunized with mouse apoB-100. The immune response against apoB-100 was robust, as judged by high titers of antibodies against mouse apoB-100. After fusing the splenic lymphocytes of the apoB-48-only mice with a myeloma cell line, we identified and cloned hybridomas that produced mouse apoB-100-specific monoclonal antibodies. Those antibodies were useful for developing sensitive and specific immunoassays for mouse apoB-100. This study illustrates the feasibility and utility of using gene-targeted mice to develop monoclonal antibodies against mouse proteins.
Zlot
C H
CH
Gladstone Institute of Cardiovascular Disease, San Francisco, CA 94141-9100, USA.
Flynn
L M
LM
Véniant
M M
MM
Kim
E
E
Raabe
M
M
McCormick
S P
SP
Ambroziak
P
P
McEvoy
L M
LM
Young
S G
SG
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D000918:Antibody Specificity; D053299:Apolipoprotein B-100; D053283:Apolipoprotein B-48; D001055:Apolipoproteins B / Q000096:biosynthesis / Q000172:deficiency / Q000235:genetics / Q000276:immunology*; D018390:Gene Targeting; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D007118:Immunoassay; D051379:Mice; D018345:Mice, Knockout
D000911:Antibodies, Monoclonal; D053299:Apolipoprotein B-100; D053283:Apolipoprotein B-48; D001055:Apolipoproteins B