Human and murine immunoglobulin expression vector cassettes

Gary R. McLean, Antonio Nakouzi, Arturo Casadevall, Nancy S. Green

Research output: Contribution to journalArticlepeer-review

Abstract

We describe the construction of new immunoglobulin (Ig) expression vectors and their use in the production of recombinant chimeric Ig molecules in transfected mammalian cells. The vectors contain the cDNA encoding the constant regions of human (μ, α1, γ1, γ2, γ3, γ4, κ) and murine (μ, γ2a, κ) Ig heavy and light chains. Unique restriction sites flanking the Ig variable region allow for replacement of variable regions generated by PCR. The CMV promoter allows for the transfection and expression of Ig in non-lymphoid cells. Distinct drug selection markers for heavy chain and light chain expression vectors allows for sequential or co-transfection of the vectors. We show that secretion of recombinant Ig can reach 1.2 μg/ml per million cells per day for transfected B cells. Replacement of the variable region results in the production of functional Ig retaining antigen specificity.

Original languageEnglish (US)
Pages (from-to)837-845
Number of pages9
JournalMolecular Immunology
Volume37
Issue number14
DOIs
StatePublished - Oct 2000
Externally publishedYes

Keywords

  • Immunoglobulin
  • Plasmid
  • Recombinant
  • Transfection
  • cDNA

ASJC Scopus subject areas

  • Immunology
  • Molecular Biology

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