Rat monoclonal antibodies to human apolipoprotein B: advantages and applications.
Eight monoclonal antibodies (Mabs) to human serum low density lipoprotein (LDL) were derived from the fusion of spleen cells, from LOU rats immunized with human LDL, and the rat myeloma line IR983F. These Mabs were characterized in terms of isotype, specificity, and affinity. Competitive experiments indicated that the epitopes that were recognized could be grouped into three patterns depending on their apparent affinity for apoB-containing lipoprotein particles such as LDL, very low density lipoproteins (VLDL), or intermediate density lipoproteins (IDL). Six epitopes have been mapped in relation to elements of the sequence of apolipoprotein B-100 (apoB-100) and some have been assigned to the middle part of the median thrombolytic fragment T3, a region not yet well targeted by mouse Mabs. The presence of lipids for the expression of the epitopes was studied and confirmed a lipid dependence for epitopes that are close to the T2/T3 cleavage site. The capacity of binding to the LDL receptor was also tested; among the Mabs we described, one inhibited the uptake and degradation of LDL to HeLa cells receptor. Finally, some antibodies were able to precipitate LDL in gel.
Fievet
C
C
INSERM U 279 et SERLIA, Institut Pasteur, Lille, France.
Durieux
C
C
Milne
R
R
Delaunay
T
T
Agnani
G
G
Bazin
H
H
Marcel
Y
Y
Fruchart
J C
JC
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology*; D000915:Antibody Affinity; D000918:Antibody Specificity; D001055:Apolipoproteins B / Q000276:immunology*; D015153:Blotting, Western; D000939:Epitopes / Q000276:immunology; D005260:Female; D006801:Humans; D006825:Hybridomas; D008077:Lipoproteins, LDL / Q000276:immunology / Q000378:metabolism; D010449:Peptide Mapping; D011233:Precipitin Tests; D011863:Radioimmunoassay; D051381:Rats; D011973:Receptors, LDL / Q000378:metabolism
D000911:Antibodies, Monoclonal; D001055:Apolipoproteins B; D000939:Epitopes; D008077:Lipoproteins, LDL; D011973:Receptors, LDL