Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving antigen-antibody binding – specific binding protein...
Patent
1997-11-26
1999-07-06
Feisee, Lila
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving antigen-antibody binding, specific binding protein...
435 71, 435 701, 435 691, 435 703, 4353201, 4352523, 435325, 530350, 536 235, G01N 3353, C12N 1500, C07H 2104
Patent
active
059196360
DESCRIPTION:
BRIEF SUMMARY
FIELD OF THE INVENTION
This invention relates generally to cell lines used in the recombinant production, screening or measurement of protein or protein interactions in vitro.
BACKGROUND OF THE INVENTION
The primary human embryonic kidney (HEK) 293 cell line is a permanent line of cells transformed by sheared human adenovirus type 5 (Ad 5) DNA. The cells are particularly sensitive to human adenovirus, are highly permissive for adenovirus DNA, and contain and express the transforming genes of Ad5. This is a hypotriploid human cell line. See, F. Graham et al., J. Gen. Virol., 36:59-72 (1977); T. Harrison et al., Virology, 77:319-329 (1977).
This cell line, which is readily available from commercial sources, such as the American Type Culture Collection, is used extensively in in vitro assays, and for the production of recombinant proteins and viruses. However, in washing steps which are conventionally and repeatedly employed in such in vitro assays and other manipulations of these cells, the cells readily detach or are washed away from the plates or dishes in which the studies are performed. This problem typically results in inaccurate, unreliably low measurement or collection of the protein, peptide or interaction to which the assay is directed.
There remains a need in the art for a cell substrate useful in in vitro manipulations in genetic engineering, which permits the measurement of accurate results.
BRIEF SUMMARY OF THE INVENTION
In one aspect, the invention provides improved HEK 293 cells, which cells are 293 cells which have been transfected with a mammalian macrophage scavenger receptor gene. Preferably, this gene is the human Type I or II
In another aspect, the invention provides a method of enhancing the ability of HEK 293 cells to attach in tissue culture. This method involves the steps of transfecting 293 cells with a selected mammalian macrophage scavenger receptor gene.
In yet another aspect, the invention provides a method of screening compounds for biological activity which involves screening the improved 293 cells of the invention. In this method, the improved 293 cells have been further transfected with a selected gene and are then screened for expression of the selected gene. The cells expressing the selected genes are incubated in the presence of a compound of unknown biological activity, and then screened for the ability of the compound to affect the expressed gene product or its function.
Other aspects and advantages of the present invention are described further in the following detailed description of the preferred embodiments thereof .
BRIEF DESCRIPTION OF THE DRAWINGS
ID NO:2! sequences of the human macrophage scavenger receptor type I. ID NO:4! sequences of the human macrophage scavenger receptor type II.
DETAILED DESCRIPTION OF THE INVENTION
The present invention provides an improved human embryonic kidney cell line, 293. The inventors have surprisingly found that human embryonic kidney (HEK) 293 cells transfected with a mammalian macrophage scavenger receptor gene demonstrate an enhanced ability to attach to a solid support as compared to conventional, unmodified 293 cells. In contrast to unmodified 293 cells, the improved 293 cells of the invention are not as readily washed away as unmodified 293 cells under the normal conditions of biological assays. Thus, the improved 293 cells of the invention are particularly well suited for use in in vitro studies and other applications for which unmodified 293 cells may be used.
As used herein "solid support" is any surface used for culturing, for in vitro assays, and the like. For example, a typical solid support is a plastic tissue culture plate, or a multi-well plate, hollow fibers, a test tube, conventionally employed plastic beads, glass beads, etc. Other solid supports are well known to those of skill in the art.
By "enhanced ability to attach" is meant that the transfected cells of this invention attach to the solid support with sufficient avidity to resist detachment which normally occurs with untransfected 293 cells ca
REFERENCES:
patent: 5510466 (1996-04-01), Krieger et al.
Matsumoto, et. al., Human macrophage scavenger receptors: Primary structure, expression, and localization in atherosclerotic lesions. Proc. Natl. Acad. Sci., Dec. (1990) 87; 9133-9137.
Sprengel, et. al., Molecular Cloning and Expression of cDNA Encoding a Peripheral-type Benzodiazepine Receptor. Journal of Biological Chemistry, Dec. 5 (1989) 264; No. 34; 20145-20421.
Kodama, et. al., Type I macrophage scavanger receptor contains .alpha.-helical and collagen-like coiled coils. Nature. Feb. 8 (1990) 343; 531-535.
Brawner Mary Ellen
Elshourbagy Nabil Abd Elsalam
Lysko Paul George
Basi Nirmal S.
Eagle Alissa M.
Feisee Lila
Hecht Elizabeth J.
King William T.
LandOfFree
Attachment enhanced 293 cells does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Attachment enhanced 293 cells, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Attachment enhanced 293 cells will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-898323