The generation of monoclonal antibodies against human peroxisome proliferator-activated receptors (PPARs).
J Atheroscler Thromb
; 9(5): 233-42, 2002.
Article
en En
| MEDLINE
| ID: mdl-12409633
ABSTRACT
Monoclonal antibodies (Mabs) are valuable reagents for the purification, characterization and immunolocalization of proteins. In this study, we raised Mabs against human peroxisome proliferator-activated receptors (PPARs) using baculovirus particles displaying surface glycoprotein gp64-fusion proteins as the immunizing agent. In this system, to display fusion proteins on the viral surface, the amino terminal sequences of human PPARd and PPARg2 are inserted in-frame between the signal sequence and the mature domain of the gp64 nucleotide sequence.Mabs were raised by immunization with whole virus without a purification of the target antigens. The Mabs generated by this novel method were shown to recognize not only the gp64-PPARs fusion protein, but also mature, expressed proteins by a wide variety of techniques, including immunohistochemistry, immunoblotting, and electrophoretic mobility shift assays (EMSAs). Transfection of the transfer vector containing a nucleotide sequence encoding less than 30 amino acids along with linearized baculovirus DNA allows for the production of a high affinity antibody against the corresponding mature form. This method is of potential utility in that it allows the production of valuable antibodies without the requirement of a protein purification step.
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Banco de datos:
MEDLINE
Asunto principal:
Factores de Transcripción
/
Receptores Citoplasmáticos y Nucleares
/
Anticuerpos Monoclonales
Límite:
Animals
/
Female
/
Humans
Idioma:
En
Año:
2002
Tipo del documento:
Article