Antibody-producing cells

[siRNA]

There are several methods to generate antibody-producing cell lines. These methods are aimed to deliver engineered DNA in eukaryotic cell in a stable expression manner (process known as "stable transfection"). Various materials and compounds are used in the process of transfection, including polymers, liposomes and nanoparticles. Among the most unreliable ways of transfection is a method in where calcium phosphate is used. Other methods use high organic mixes, dendrimers, cationic polymers, DEAE-dextran, polyethylenimine, electroporation, heat shock, and magnetofection.

For the majority of transfection applications, it is enough if the transfected gene is only transitive expressed, however to generate a stable cell line - transfected gene should remain in the genome of the cells and the daughter’s cells, stabilised transfection should occur. For the proper performance of this process, another gene should be co-transfected, which will give to the cell some advantage, such as a resistance to some toxins (usually antibiotic resistance genes are used). If the toxin to which the gene is co-transfected offers resistance (antibiotic-resistant) and then is added to the cells - only those cells encoding resistance genes will be able to proliferate, while other cells will die. The general substance for stabilised transfection is Geneticin, or the so-called G418, which will be the toxin, that can be neutralised by the persistent product of gene neomycin.

Overall, generation of antibody-producing cell line takes 2-4 months with proper cell line generation, analysis, and characterization. Altogen (http://www.altogenservices.com) offer commercial services of stable cell line generation.