TY - JOUR
T1 - Genetically engineered intracellular single-chain antibodies in gene therapy
AU - Bilbao, Guadalupe
AU - Contreras, Juan Luis
AU - Curiel, David T.
PY - 2002/10
Y1 - 2002/10
N2 - 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.
AB - 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.
KW - Colony lift assay
KW - Gene therapy
KW - Heavy chain (VH)
KW - Intracellular single-chain antibody (scFv)
KW - Light chain (VL)
UR - http://www.scopus.com/inward/record.url?scp=0036798045&partnerID=8YFLogxK
U2 - 10.1385/MB:22:2:191
DO - 10.1385/MB:22:2:191
M3 - Article
C2 - 12405266
AN - SCOPUS:0036798045
VL - 22
SP - 191
EP - 211
JO - Applied Biochemistry and Biotechnology - Part B Molecular Biotechnology
JF - Applied Biochemistry and Biotechnology - Part B Molecular Biotechnology
SN - 1073-6085
IS - 2
ER -