Gene-based vaccination and screening methods to develop monoclonal antibodies

M. Pirouz Daftarian, Aram Vosoughi, Vance Lemmon

Research output: Chapter in Book/Report/Conference proceedingChapter

1 Scopus citations

Abstract

Gene-based in vivo electroporation has the potential to be used as a "protein-free" method to elicit immune responses and to generate monoclonal antibodies (mAb) against proteins/peptides in hosts. However, the method is very useful to raise mAbs against proteins and peptides and not for carbohydrates, lipids, or haptens. Nevertheless, making mAb using this potent method faces a few challenges: the parameters of the electroporation needs further standardized, the final boost still needs protein antigens, and the primary screening of the clones requires purified protein. We present methods to overcome these challenges by an optimized electroporation framework and a method to use transiently transfected cells for the final boost, as well as for screening of the resulting clones via the use of an "In-Cell Western" method.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages337-346
Number of pages10
Volume1121
ISBN (Print)9781461496311
DOIs
StatePublished - 2014

Publication series

NameMethods in Molecular Biology
Volume1121
ISSN (Print)10643745

Keywords

  • Antibodies electroporation
  • Gene
  • In-Cell Western
  • Screening
  • Vaccine

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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    Daftarian, M. P., Vosoughi, A., & Lemmon, V. (2014). Gene-based vaccination and screening methods to develop monoclonal antibodies. In Methods in Molecular Biology (Vol. 1121, pp. 337-346). (Methods in Molecular Biology; Vol. 1121). Humana Press Inc.. https://doi.org/10.1007/978-1-4614-9632-8_30