Generation of Antibody-producing Hybridomas Following One Single Immunization with a Targeted DNA Vaccine
Overview
Authors
Affiliations
The standard protocol for generating antibody (Ab)-producing hybridomas is based on fusion of plasmacytoma cells with Ab-producing B cells harvested from immunized mice. To increase the yield of hybridomas, it is important to use immunization protocols that induce a high frequency of B cells producing specific Abs. Our laboratory has developed a vaccine format, denoted vaccibody that promotes the immune responses towards the delivered antigen. The vaccine format targets antigens in a bivalent form to surface receptors on antigen-presenting cells (APCs). Here, we used the fluorescent protein (FP) mCherry as antigen and targeted it to APCs by use of either the natural ligand CCL3/MIP-1α or single-chain variable fragment specific for major histocompatibility complex class II. The vaccine format was delivered to mouse muscle as DNA combined with electroporation. By this procedure, we developed two monoclonal Abs that can be utilized to detect the FC mCherry in various applications. The data suggest that the targeted DNA vaccine format can be utilized to enhance the number of Ab-producing hybridomas and thereby be a tool to improve the B cell hybridoma technology.
Braathen R, Spang H, Hinke D, Blazevski J, Bobic S, Fossum E Mol Ther Methods Clin Dev. 2020; 17:378-392.
PMID: 32128342 PMC: 7044496. DOI: 10.1016/j.omtm.2020.01.007.
Lysen A, Braathen R, Gudjonsson A, Tesfaye D, Bogen B, Fossum E Sci Rep. 2019; 9(1):1820.
PMID: 30755656 PMC: 6372594. DOI: 10.1038/s41598-018-38080-7.
Zhou P, Chionh Y, Irac S, Ahn M, Jia Ng J, Fossum E Sci Rep. 2016; 6:38597.
PMID: 27934903 PMC: 5146944. DOI: 10.1038/srep38597.
DNA immunization as a technology platform for monoclonal antibody induction.
Liu S, Wang S, Lu S Emerg Microbes Infect. 2016; 5:e33.
PMID: 27048742 PMC: 4855071. DOI: 10.1038/emi.2016.27.
Kuczkowska K, Mathiesen G, Eijsink V, Oynebraten I Microb Cell Fact. 2015; 14:169.
PMID: 26494531 PMC: 4618854. DOI: 10.1186/s12934-015-0360-z.