PharmPK Discussion - Development of a cell-based assay for drug screening

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On 4 Nov 1998 at 11:45:13, Tan-yar Ling (lingty.aaa.mcmaster.ca) sent the message


Has anyone come across any information i.e. journals, publication etc...
regarding protocol for the production of a stable cell-line expressing the
human Eta receptor.

I need to develop a protocol using the endothelin receptor area.

Thanks

Tan-yar Ling

Biology-pharmacology
McMaster University

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On 6 Nov 1998 at 15:54:09, David_Bourne (david.-at-.boomer.org) sent the message


This reference is to the method used by Zeneca for ETa receptor
expression..

Needham, M., Gooding, C., Hudson, K., Antoniou, M., Grosveld, F. and
Hollis, M:  LCR/MEL: A versatile system for high level expression of
heterologous proteins in erythroid cells.  Nucl. Acids Res.  20:
997-1003, 1992.

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On 20 Nov 1998 at 09:45:37, Tan-yar Ling (lingty.at.mcmaster.ca) sent the message


Hi!  Thank-you for the reference, but there are some changes that make
that reference irrelevant.

The project to develop a protocol for the production of a stable cell
line
expressing human ETa receptor.  One of the constraints of the project is

that the stable cell line must be created by transfection of a cell line

(not expressing the ETa receptor) with pCEP4 plasmid (which has the
genes that code for the production of ETa receptor).  We have been given

the cloned human ETa receptor in pCEP4 plasmid, but need suggestions for

a cell line that we can successfully transfect.

Can you provide any suggestions as to the protocol that we might be able

to utilize?
We will be using this cell line to develop a protocol to validate the
usefulness of the cell line in High Throughput Screening for novel ETa
agonists and antagonists.

Any help or direction would be much appreciated

Thank-you

Tan-yar Ling

Honours Biology and Pharmacology Programme
McMaster University
Hamilton, Ontario

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