Monoclonal antibodies to human apolipoproteins: application to the study of high density lipoprotein subpopulations.
We produced, selected and cloned hybridomas that secrete monoclonal antibodies against human apolipoprotein (apo) A-I. All of the antibodies corresponded to the IgG(1) subclass and were named 1C11, 2B4, 2C10, 7C5, 8A4 and 8A5. The antibodies were characterized by their reactivity with whole lipoproteins, apolipoproteins, synthetic peptides and fragments generated by cleavage of the apo A-I. Three of the monoclonal antibodies studied (2B4, 2C10 and 7C5) were similarly inhibited by an amino-terminal peptide (amino acid sequence 1-20) of apo A-I, whereas antibodies 1C11, 8A4 and 8A5 had no reaction. Other results show that monoclonal antibody 1C11 recognizes an epitope located between amino acids 135-148. We evaluated the monoclonal antibody 8A4 against different HDL subpopulations by competitive displacement analysis and it showed a similar reactivity with the HDL particles: LpA-I and LpA-I:A-II. This antibody was used to standardize a sandwich ELISA to quantitate LpA-I in plasma. We conclude that these monoclonal antibodies are relevant for the study of apo A-I epitope expression and for quantitating apo A-I containing lipoparticles.
Bustos
P
P
Departamento Bioquímica Clínica e Inmunología, Facultad de Farmacia, Universidad de Concepción, Concepción, Chile. pbustos@mail.udec.cl
Ulloa
N
N
Calvo
C
C
Muller
D
D
Durán
D
D
Martínez
J
J
Salazar
L
L
Quiroga
A
A
D000818:Animals; D000911:Antibodies, Monoclonal; D001053:Apolipoproteins / Q000032:analysis* / Q000276:immunology*; D001667:Binding, Competitive; D015153:Blotting, Western; D002478:Cells, Cultured; D003488:Cyanogen Bromide; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007202:Indicators and Reagents; D017270:Lipoprotein(a) / Q000032:analysis; D008075:Lipoproteins, HDL / Q000032:analysis*; D008081:Liposomes / Q000737:chemistry; D051379:Mice; D008807:Mice, Inbred BALB C / Q000276:immunology; D013154:Spleen / Q000166:cytology
D000911:Antibodies, Monoclonal; D001053:Apolipoproteins; D007202:Indicators and Reagents; D017270:Lipoprotein(a); D008075:Lipoproteins, HDL; D008081:Liposomes; D003488:Cyanogen Bromide