Factors that affect the efficiency of cell transfection by immunoporation

Immunoporation is a recently discovered method that is able to transfect various human cell lines efficiently by targeting the cell surface antigens with antibody-coated beads. For this particular study, HL60, a cell line difficult to transfect by other methods, was used as a model to define the var...

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Bibliographic Details
Published in:Analytical biochemistry 2004-05, Vol.328 (2), p.219-224
Main Authors: Tzavelas, C, Bildirici, L, Rickwood, D
Format: Article
Language:English
Subjects:
Antigen-Antibody Reactions
Antigens, Surface - analysis
Antigens, Surface - metabolism
beta-Galactosidase - metabolism
CD antigens
Cell Line
Cell membrane
Cell Survival
Efficiency
Electroporation - methods
Fluorescent Antibody Technique - methods
HL-60 Cells
Humans
Immunoporation
K562 Cells
Time Factors
Transfection
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Summary:Immunoporation is a recently discovered method that is able to transfect various human cell lines efficiently by targeting the cell surface antigens with antibody-coated beads. For this particular study, HL60, a cell line difficult to transfect by other methods, was used as a model to define the various parameters of the cell membrane that determine the efficiency of this method. The level of antigen expression on the cell surface was the first parameter to be analyzed and experiments indicated that there is a close correlation between the level of expression of surface antigens and the efficiency of immunoporation. The mixing speed, the bead to cell ratio, and the mixing time were all found to affect the ability of the antigen-coated beads to pull holes in the cells and it was found that for HL60 cells the optimum mixing speed was 40 rpm and the bead to cell ratio was 20:1 using a mixing time of 6 h.
ISSN:0003-2697
1096-0309
DOI:10.1016/j.ab.2004.01.036