Mouse monoclonal antipeptide antibodies specific for cholesteryl ester transfer protein (CETP).
A synthetic peptide whose amino acid sequence corresponds to residues 131-142 of human cholesteryl ester transfer protein (CETP) was used as an immunogen to generate a panel of monoclonal antibodies (MAbs) specific for the intact CETP molecule. Spleen cells from BALB/c mice immunized with the peptide conjugated with keyhole limpet hemocyanin (KLH) were fused with SP2/0 myeloma cells. Two MAbs that bound fixed peptide in an enzyme-linked immunoabsorbent assay (ELISA) were partially characterized regarding their specificity and biological activity. ATM192 of the IgG1 subclass and J16-14 of the IgG3 subclass were used in a Western blot assay as well as in the ELISA. We have also shown through the use of immunoprecipitation that ATM192 can remove CETP enzyme activity from human serum without destroying the enzyme's activity. We have also shown that the antibodies can bind CETP from rabbits. The specificity studies and the lack of inhibition of enzymatic activity suggest that the MAbs bind a structural area of the CETP molecule not a part of the active binding site of the enzyme. We conclude that these antibodies can be valuable as tools for studying CETP levels in human serum as well as in tissue homogenates from rabbits and humans.
Department of Microbiology and Immunology, East Carolina University School of Medicine, Greenville, North Carolina 27858-4354, USA.
A P
AP
Thomas
A M
AM
Smith
R I
RI
Cumming
C
C
Jones
R C
RC
Thomas
K T
KT
Pleasants
H
H
Barakat
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry*; D000918:Antibody Specificity; D002352:Carrier Proteins / Q000276:immunology*; D053480:Cholesterol Ester Transfer Proteins; D002788:Cholesterol Esters / Q000737:chemistry / Q000276:immunology*; D005260:Female; D006023:Glycoproteins; D006825:Hybridomas / Q000737:chemistry; D051379:Mice; D008807:Mice, Inbred BALB C; D010455:Peptides / Q000276:immunology*
D000911:Antibodies, Monoclonal; C506642:CETP protein, human; D002352:Carrier Proteins; D053480:Cholesterol Ester Transfer Proteins; D002788:Cholesterol Esters; D006023:Glycoproteins; D010455:Peptides