title: Optimization of Gene Transfection in Murine Myeloma Cell Lines using Different Transfection Reagents.
abstract: Purification and isolation of cellular target proteins for monoclonal antibody (MAb) production is a difficult and time-consuming process. Immunization of mice with murine cell lines stably transfected with genes coding for xenogenic target molecules is an alternative method for mouse immunization and MAb production. Here we present data on transfection efficiency of some commercial reagents used for transfection of murine myeloma cell lines. Little is known about transfectability of murine myeloma cell lines by different transfection reagents. Mouse myeloma cell lines (SP2/0, NS0, NS1, Ag8, and P3U1) were transfected with pEGFP-N1 vector using Lipofectamine 2000, jetPEI and LyoVec commercial transfection reagents in different combinations. The transfection permissible HEK293-FT cell line was used as a control in transfection procedure. Transfected cells, expressing the Enhanced Green Fluorescent Protein (EGFP), were analyzed by flow cytometry 48 hrs post transfection. Our results showed transfection efficiency of 71%, 57% and 22% for HEK293-FT, 5.5%, 3.4% and 1% for SP2/0, 55.7%, 21.1% and 9.3% for NS0, 8.2%, 6% and 5.5% for NS1, 22%, 49.2% and 5.5% for Ag8 and 6.3%, 21.5% and 4.6% for P3U1 cell lines after transfection with Lipofectamine 2000, jetPEI and LyoVec reagents, respectively. Our data indicate that NS0 and Ag8 are efficiently transfected by Lipofectamine 2000 and jetPEI reagents. Finally, we propose Ag8 and NS0 cell lines as suitable host cells for efficient expression of target genes which can be used for mouse immunization and MAb production.
authors: Department of Immunology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran ; Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR, Tehran, Iran.
authors: Mahdi
authors: M
authors: Shabani
authors: Sheyda
authors: S
authors: Hemmati
authors: Reza
authors: R
authors: Hadavi
authors: Zahra
authors: Z
authors: Amirghofran
authors: Mahmood
authors: M
authors: Jeddi-Tehrani
authors: Hodjatallah
authors: H
authors: Rabbani
authors: Fazel
authors: F
authors: Shokri
keywords: Enhanced green fluorescent protein; Flow cytometry; Monoclonal antibody; Myeloma cell lines; Transfection