Intracellular and cell surface displayed single-chain diabodies

Kontermann, Roland E.; Müller, Rolf

doi:10.1016/s0022-1759(99)00062-9

Cited by 39 publications

(16 citation statements)

References 41 publications

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“…Based on these assays, a fusion protein of the PDGF receptor transmembrane domain 14 with a ␤-glucuronidase molecule that was C-terminally truncated at Thr-633 (␤633-PDGFR) turned out to be the most promising construct. This C-terminal truncation was essential because of Cterminal processing of the de novo synthesized polypeptide.…”

Section: Resultsmentioning

confidence: 99%

Cell surface display of a lysosomal enzyme for extracellular gene-directed enzyme prodrug therapy

2001

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Prodrug conversion is a promising approach to cytotoxic gene therapy if an efficient transfer of the generated drug to adjacent cells can be achieved. To maximize the efficacy of this strategy we sought to develop a system that is based on a human enzyme, acts extracellularly yet in close vicinity of the transduced cell and can be used with multiple prodrugs. Results obtained with a secreted version of human ␤-glucuronidase suggested that this enzyme could be a suitable candidate, although a more stringent retention of the

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Section: Resultsmentioning

confidence: 99%

Cell surface display of a lysosomal enzyme for extracellular gene-directed enzyme prodrug therapy

2001

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“…[18,19] This construction facilitates chain pairing and minimizes the refolding and aggregation problems encountered when the two chains are expressed individually. [18,20,21] Importantly, the affinity and stability of scFvs with the (GGGGS) 3 linker are similar to those of the native antibody, even though the scFvs are more specific. [22] As described in many studies, two rounds of panning were sufficient to enrich for the desired sequences.…”

Section: Discussionmentioning

confidence: 99%

Construction, expression and characterisation of a single chain variable fragment in the Escherichia coli periplasmic that recognise MCF-7 breast cancer cell line

Mahgoub

Bolad

2014

J Can Res Ther

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“…The advances in antibody cloning technology have greatly facilitated the genetic manipulation of antibody fragments and permitted the development of a large variety of engineered antibody molecules for research, diagnosis and therapy (Boss et al, 1984;Kontermann and Muller, 1999). It is possible for the cloned antibodies to have improved the affinity and specificity of antigen binding by mimicking somatic hypermutation during an immune response (Gram et al, 1992).…”

Section: Introductionmentioning

confidence: 99%

Construction of single-chain variable fragment antibodies against MCF-7 breast cancer cells

Zuhaida¹,

Ali²,

Tamilselvan³

et al. 2013

Genet. Mol. Res.

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ABSTRACT.A phage display library of single chain variable fragment (scFv) against MCF-7 breast cancer cells was constructed from C3A8 hybridoma cells. RNA from the C3A8 was isolated, cDNA was constructed, and variable heavy and light immunoglobulin chain gene region were amplified using PCR. The variable heavy and light chain gene regions were combined with flexible linker, linked to a pCANTAB 5E phagemid vector and electrophoresed into supE strain of Escherichia coli TG1 cells. Forty-eight clones demonstrated positive binding activity to MCF-7 breast cancer cell membrane fragments and the strongest of 48 clones was selected for analysis. The anti-MCF-7 library evaluated by SfiI and NotI digests demonstrated that anti-MCF-7 scFv antibodies possess individual patterns that should be able to recognize distinct human breast cancer cells. The C3A8 scFv, with an apparent molecular weight of 32 kDa, showed high homology (99%) with single chain antibody against rice stripe virus protein P20. In summary, the anti MCF-7 scFv antibody can be used for pretargeting breast cancer for clinical diagnosis of patients; it also has potential for therapeutic applications.

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Intracellular and cell surface displayed single-chain diabodies

Cited by 39 publications

References 41 publications

Cell surface display of a lysosomal enzyme for extracellular gene-directed enzyme prodrug therapy

Cell surface display of a lysosomal enzyme for extracellular gene-directed enzyme prodrug therapy

Construction, expression and characterisation of a single chain variable fragment in the Escherichia coli periplasmic that recognise MCF-7 breast cancer cell line

Construction of single-chain variable fragment antibodies against MCF-7 breast cancer cells

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