Method to select and transfect cell subpopulations

Details Method to select and transfect cell subpopulations Method to select and transfect cell subpopulations

1999-01-27

2001-05-08

Schwartzman, Robert A. (Department: 1635)

Chemistry: molecular biology and microbiology

Treatment of micro-organisms or enzymes with electrical or...

Concentration, separation, or purification of micro-organisms

C435S459000

Reexamination Certificate

active

06228624

ABSTRACT:

BACKGROUND OF THE INVENTION
The infection or transfection of cells by infectious viruses or retroviruses has been known for many years. Many methods for improving the rate of transfection have been used. Calcium chloride treatment of the cells has been used to allow naked DNA to pass through the cellular membrane. Improved vectors have been developed for increasing the level of infection. The use of lipids or liposomes to deliver the genetic material have also been used with some amount of success.
Particle mediated delivery has been used to transform numerous types of cells with a gold microparticle mediated delivery of DNA. The technique has been described in numerous publications and has advanced enough to merit a product which has been marketed by BioRad under the name Biolistic PDS. The use of this and similar systems are described in U.S. Pat. Nos. 4,945,050; 5,015,580; and 5,120,657 and international patent applications WO 95/29703 and WO 93/17706. An improvement upon the particle mediated delivery was made by Palsson, as disclosed in U.S. Pat. No. 5,534,423. The Palsson method includes loading a vector and the cells to be transformed into a centrifuge apparatus and spinning it to bring the vectors into contact with the cells. A spinnable disk and an electrolytic system are also envisioned.
SUMMARY OF THE INVENTION
The instant invention provides a method for transfection of target cells by which the transfection efficiency of vectors and target cells is improved by bringing the target cell and the vector in contact. This is achieved by immobilization of both the vector containing the relevant genetic material and the ligand specific for the target cells on a particle. When such particle is now incubated with a test sample consisting of a cell mixture containing the target cells, the ligand specific for the target cells will bind to the target cells and thus bring the vector in contact with the target cells. The target cells can now be separated from the non target cells by isolation of the particle-cell construct, thus achieving both enrichment of target cells and transfection of the target cells.


REFERENCES:
patent: 5460831 (1995-10-01), Kossovsky et al.
patent: 9423738 (1994-10-01), None
R. Padmanabhan et al., “Purification of Transiently Transfected Cells by Magnetic Affinity Cell Sorting”, Analytical Biochemistry, 170: 341-348 (1988).
M. Pilon et al., “Transient Expression of the CD2 Cell Surface Antigen as a Sortable Marker to Monitor High Frequency Transfection of Human Primary B Cells”, The Journal of Immunology, 146: 1047-1051 (1991).
T. Giordano et al., “Isolation of a Population of Transiently Transfected Quiescent and Senescent Cells by Magnetic Affinity Cell Sorting”, Experimental Cell Research, 192: 193-197 (1991).

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