» Articles » PMID: 21955209

Generation of Antibody-producing Hybridomas Following One Single Immunization with a Targeted DNA Vaccine

Overview
Journal Scand J Immunol
Date 2011 Sep 30
PMID 21955209
Citations 8
Authors
Affiliations
Soon will be listed here.
Abstract

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.

Citing Articles

A DNA Vaccine That Encodes an Antigen-Presenting Cell-Specific Heterodimeric Protein Protects against Cancer and Influenza.

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.


Dendritic cell targeted Ccl3- and Xcl1-fusion DNA vaccines differ in induced immune responses and optimal delivery site.

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.


Unlocking bat immunology: establishment of Pteropus alecto bone marrow-derived dendritic cells and macrophages.

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.


Lactobacillus plantarum displaying CCL3 chemokine in fusion with HIV-1 Gag derived antigen causes increased recruitment of T cells.

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.


References
1.
Kohler G, Milstein C . Continuous cultures of fused cells secreting antibody of predefined specificity. 1975. J Immunol. 2005; 174(5):2453-5. View

2.
Di Cristina M, Nunziangeli L, Giubilei M, Capuccini B, dEpiscopo L, Mazzoleni G . An antigen microarray immunoassay for multiplex screening of mouse monoclonal antibodies. Nat Protoc. 2010; 5(12):1932-44. DOI: 10.1038/nprot.2010.161. View

3.
Mir L, Bureau M, Gehl J, Rangara R, Rouy D, CAILLAUD J . High-efficiency gene transfer into skeletal muscle mediated by electric pulses. Proc Natl Acad Sci U S A. 1999; 96(8):4262-7. PMC: 16320. DOI: 10.1073/pnas.96.8.4262. View

4.
Barber B . The immunotargeting approach to adjuvant-independent subunit vaccine design. Semin Immunol. 1997; 9(5):293-301. DOI: 10.1006/smim.1997.0085. View

5.
Larsson K, Wester K, Nilsson P, Uhlen M, Hober S, Wernerus H . Multiplexed PrEST immunization for high-throughput affinity proteomics. J Immunol Methods. 2006; 315(1-2):110-20. DOI: 10.1016/j.jim.2006.07.014. View