Unknown

Dataset Information

0

Isolation of a high-affinity stable single-chain Fv specific for mesothelin from DNA-immunized mice by phage display and construction of a recombinant immunotoxin with anti-tumor activity.


ABSTRACT: Mesothelin is a differentiation antigen present on the surface of ovarian cancers, mesotheliomas, and several other types of human cancers. Because among normal tissues, mesothelin is present only on mesothelial cells, it represents a good target for antibody-mediated delivery of cytotoxic agents. In the present study mice were immunized with an eukaryotic expression vector coding for mesothelin. When high serum antibody titers were obtained, a phage display library was made from the splenic mRNA of these mice. After three rounds of panning on recombinant mesothelin, a single-chain Fv (scFv)-displaying phage was selected that bound specifically to recombinant mesothelin and mesothelin-positive cells. The scFv was used to construct an immunotoxin by genetically fusing it with a truncated mutant of Pseudomonas exotoxin A. The purified immunotoxin binds mesothelin with high affinity (Kd 11 nm), is stable for over 40 hr at 37 degrees C and is very cytotoxic to cells expressing mesothelin. It also produces regressions of tumors expressing mesothelin. This combination of selective cytotoxicity, high activity, and stability makes the immunotoxin a good candidate for development as a therapeutic agent. This work also shows that DNA immunization can be used to isolate and clone antibodies against epitopes present on human proteins in their native conformation.

SUBMITTER: Chowdhury PS 

PROVIDER: S-EPMC18478 | biostudies-literature | 1998 Jan

REPOSITORIES: biostudies-literature

altmetric image

Publications

Isolation of a high-affinity stable single-chain Fv specific for mesothelin from DNA-immunized mice by phage display and construction of a recombinant immunotoxin with anti-tumor activity.

Chowdhury P S PS   Viner J L JL   Beers R R   Pastan I I  

Proceedings of the National Academy of Sciences of the United States of America 19980101 2


Mesothelin is a differentiation antigen present on the surface of ovarian cancers, mesotheliomas, and several other types of human cancers. Because among normal tissues, mesothelin is present only on mesothelial cells, it represents a good target for antibody-mediated delivery of cytotoxic agents. In the present study mice were immunized with an eukaryotic expression vector coding for mesothelin. When high serum antibody titers were obtained, a phage display library was made from the splenic mRN  ...[more]

Similar Datasets

| S-EPMC8317202 | biostudies-literature
| S-EPMC5737338 | biostudies-literature
| S-EPMC2630973 | biostudies-literature
| S-EPMC45374 | biostudies-other
| S-EPMC5058652 | biostudies-literature
| S-EPMC406508 | biostudies-literature
| S-EPMC4674367 | biostudies-literature
| S-EPMC3127757 | biostudies-literature
| S-EPMC3785514 | biostudies-literature
| S-EPMC2362362 | biostudies-other