Genetically engineered intracellular single-chain antibodies in gene therapy

Guadalupe Bilbao, Juan Luis Contreras, David T. Curiel

Research output: Contribution to journalArticle

9 Scopus citations

Abstract

The delineation of the molecular basis of cancer allows for the possibility of specific intervention at the molecular level for therapeutic purposes. To a large extent, the genetic lesions associated with malignant transformation and progression are being identified. Thus, not only in the context of inherited genetic diseases, but also for many acquired disorders, characteristic aberrancies of patterns of gene expression may be precisely defined. It is therefore clear that elucidation of the genetic basis of inherited and acquired diseases has rendered gene therapy both a novel and rational approach for these disorders. To this end, three main strategies have been developed: mutation compensation, molecular chemotherapy, and genetic immuno-potentiation. Mutation compensation relies on strategies to ablate activated oncogenes at the level of DNA (triplex), messenger RNA (antisense or ribozyme), or protein (intracellular single-chain antibodies), and augment tumor suppressor gene expression. This article will review in detail practical procedures to generate a single-chain intracellular antibody (scFv). We will emphasize in this article the different steps in our protocol that we have employed to develop scFvs to a variety of target proteins.

Original languageEnglish
Pages (from-to)191-211
Number of pages21
JournalApplied Biochemistry and Biotechnology - Part B Molecular Biotechnology
Volume22
Issue number2
DOIs
StatePublished - Oct 2002
Externally publishedYes

Keywords

  • Colony lift assay
  • Gene therapy
  • Heavy chain (VH)
  • Intracellular single-chain antibody (scFv)
  • Light chain (VL)

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