Detection of electronegative low density lipoprotein (LDL-) in plasma and atherosclerotic lesions by monoclonal antibody-based immunoassays.
OBJECTIVE
To produce a monoclonal antibody (MAb) against electronegative LDL (LDL-) for detecting this modified lipoprotein in blood plasma and tissues.


METHODS
LDL- was isolated from human blood plasma and used as an antigen for immunization of Balb/c mice. Lymphocytes of immunized mice were fused with myeloma cells (SP2/0) to obtain the hybridomas. LDL- was detected in blood plasma and atherosclerotic lesions of humans and rabbits by MAb-based ELISA and immunohistochemistry, respectively.


RESULTS
LDL- concentrations were higher (P < 0.05) in the blood plasma of hypercholesterolemic subjects (HC, 248 +/- 77 mg/dL of total cholesterol) than in normolipidemic subjects (NL, 173 +/- 82 mg/dL of total cholesterol) and rabbits (HC, 250 +/- 15 mg/dL of cholesterol versus NL, 81 +/- 12 mg/dL of cholesterol). Moreover, LDL- was detected in the atherosclerotic lesions of humans and rabbits.


CONCLUSIONS
These MAb-based immunoassays are adequate to detect LDL- in biological samples and represent an important tool for investigating the role of LDL- in atherosclerosis.
Damasceno
Nágila R T
NR
Department of Nutrition, School of Public Health, University of São Paulo, SP, Brazil.
Sevanian
Alex
A
Apolinário
Elaine
E
Oliveira
Joelma M A
JM
Fernandes
Irene
I
Abdalla
Dulcineia S P
DS
D000368:Aged; D000369:Aged, 80 and over; D000911:Antibodies, Monoclonal; D000915:Antibody Affinity; D001013:Aorta, Thoracic / Q000378:metabolism / Q000473:pathology; D050197:Atherosclerosis / Q000378:metabolism* / Q000473:pathology; D008078:Cholesterol, LDL / Q000032:analysis* / Q000097:blood / Q000276:immunology; D002852:Chromatography, Ion Exchange; D003429:Cross Reactions; D005260:Female; D006801:Humans; D007118:Immunoassay; D015151:Immunoblotting; D008297:Male; D008875:Middle Aged
D000911:Antibodies, Monoclonal; D008078:Cholesterol, LDL