We have produced and characterized six urine monoclonal antibodies to human apolipoprotein A-I named A-I-9, A-I-12, A-I-15, A-I-16, A-I-19, and A-I-57. All monoclonal antibodies were specific for apolipoprotein A-I and bound between 55% and 100% of 125I-labeled high density lipoproteins (HDL) in a fluid phase radioimmunoassay. All antibodies possessed a higher affinity to apoA-I in HDL than to free, delipidated apoA-I. Two of them, particularly A-I-12 and A-I-15, which were directed to the same or very close epitopes on the molecule, recognized very poorly the delipidated protein. Binding of apoA-I to phospholipid restored the immunoreactivity of the monoclonal antibodies ot the protein suggesting that lipids play and important role in determining the immunochemical structure of apoA-I. Using CNBr fragments and synthetic peptides, the epitopes for the antibodies were mapped as follows: A-I-19, CNBr fragment 1; A-I-12 and 15, CNBr fragment 2; A-I-9 and A-I-16, CNBr fragment 3; A-I-57, CNBr fragment 4. Antibody A-I-57 failed to recognize a mutant form of apoA-I, A-I(Milano) (Arg173 → Cys) by immunoblotting and by competitive radioimmunoassay demonstrating that substitution of a single amino acid in human apoA-I may cause the loss of an antigenic determinant.

IMMUNOCHEMICAL CHARACTERIZATION OF 6 MONOCLONAL-ANTIBODIES TO HUMAN APOLIPOPROTEIN-A-I - EPITOPE MAPPING AND EXPRESSION / S. MARCOVINA, S. FANTAPPIE, A. ZOPPO, G. FRANCESCHINI, A. CATAPANO. - In: JOURNAL OF LIPID RESEARCH. - ISSN 0022-2275. - 31:3(1990), pp. 375-384.

IMMUNOCHEMICAL CHARACTERIZATION OF 6 MONOCLONAL-ANTIBODIES TO HUMAN APOLIPOPROTEIN-A-I - EPITOPE MAPPING AND EXPRESSION

G. FRANCESCHINI
Penultimo
;
A. CATAPANO
Ultimo
1990

Abstract

We have produced and characterized six urine monoclonal antibodies to human apolipoprotein A-I named A-I-9, A-I-12, A-I-15, A-I-16, A-I-19, and A-I-57. All monoclonal antibodies were specific for apolipoprotein A-I and bound between 55% and 100% of 125I-labeled high density lipoproteins (HDL) in a fluid phase radioimmunoassay. All antibodies possessed a higher affinity to apoA-I in HDL than to free, delipidated apoA-I. Two of them, particularly A-I-12 and A-I-15, which were directed to the same or very close epitopes on the molecule, recognized very poorly the delipidated protein. Binding of apoA-I to phospholipid restored the immunoreactivity of the monoclonal antibodies ot the protein suggesting that lipids play and important role in determining the immunochemical structure of apoA-I. Using CNBr fragments and synthetic peptides, the epitopes for the antibodies were mapped as follows: A-I-19, CNBr fragment 1; A-I-12 and 15, CNBr fragment 2; A-I-9 and A-I-16, CNBr fragment 3; A-I-57, CNBr fragment 4. Antibody A-I-57 failed to recognize a mutant form of apoA-I, A-I(Milano) (Arg173 → Cys) by immunoblotting and by competitive radioimmunoassay demonstrating that substitution of a single amino acid in human apoA-I may cause the loss of an antigenic determinant.
Settore BIO/14 - Farmacologia
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2434/182483
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