The use of recombinant vaccinia virus to generate monoclonal antibodies against the cell‐surface glycoprotein endoglin

FEBS Letters

Páez, Eduardo³  Herreros, Lourdes²  Cabañas, Carlos²  Luque, Alfonso²  Raab, Ulla³  Letamendı́a, Ainhoa³  Sánchez-Madrid, Francisco¹  Bernabeu, Carmelo³ 

[1] Servicio de Inmunologı́a, Hospital de La Princesa, 28006 Madrid, Spain;Departamento de Bioquı́mica y Biologı́a Molecular III, Facultad de Medicina, Universidad Complutense, 28040 Madrid, Spain;Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Cientı́ficas (CSIC), 28006 Madrid, Spain

关键词: Monoclonal antibody;    Vaccinia virus;    Surface antigen;    Endoglin (human);   

DOI  :  10.1016/S0014-5793(97)00860-0

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Characterization of novel cell-surface protein molecules, initially identified by cDNA cloning techniques, usually requires the generation of specific antibodies to further analyze their biochemical and/or functional properties. Here we report a simple method, using recombinant vaccinia virus, for the generation of monoclonal antibodies (mAb) to the cell-surface antigen endoglin. A recombinant vaccinia virus carrying a cDNA encoding human endoglin was inserted into the thymidine kinase locus under the control of the 7.5k vaccinia virus promoter. Infection of Balb/c mice with this recombinant virus led to the generation of specific polyclonal antibodies, as demonstrated by the antisera reactivity against human endoglin transfectants. The spleen cells of these infected animals were fused to myeloma cells, allowing efficient generation of several hybridomas which secrete mAbs to human endoglin, as evidenced by their reactivity with purified endoglin as well as with endoglin transfectants. Some of the mAbs selected seem to be specific for regions of endoglin conserved among different species as evidenced by their cross-reactivity with chicken endoglin. These results underline the utility of recombinant vaccinia virus to generate antibodies with novel properties to new cell surface proteins such as endoglin.

【 授权许可】

Unknown