In vivo Production of Functional Single-Chain Fv Fragment with an N-Terminal-Specific Bio-orthogonal Reactive Group

ChemBioChem ◽  
2010 ◽  
Vol 11 (4) ◽  
pp. 498-501 ◽  
Author(s):  
Edwardraja Selvakumar ◽  
Neelamegam Rameshkumar ◽  
Sun-Gu Lee ◽  
Soo-Jae Lee ◽  
Hyung-Soon Park
Keyword(s):  
Single Chain ◽  
Reactive Group ◽  
Single Chain Fv ◽  
Single Chain Fv Fragment

Enhancing the thermal stability of a single-chain Fv fragment by in vivo global fluorination of the proline residues

2011 ◽  
Vol 7 (1) ◽  
pp. 258-265 ◽  
Author(s):  
Selvakumar Edwardraja ◽  
Sokalingam Sriram ◽  
Raghunathan Govindan ◽  
Nediljko Budisa ◽  
Sun-Gu Lee
Keyword(s):  
Thermal Stability ◽  
Single Chain ◽  
Single Chain Fv ◽  
Single Chain Fv Fragment ◽  
Thermal Stability Of

In vitro and in vivo inhibition of complement activity by a single-chain Fv fragment recognizing human C5

1995 ◽  
Vol 32 (16) ◽  
pp. 1183-1195 ◽  
Author(s):  
Mark J Evans ◽  
Scott A Rollins ◽  
Dennis W Wolff ◽  
Russell P Rother ◽  
Allen J Norin ◽  
...  
Keyword(s):  
Single Chain ◽  
Complement Activity ◽  
Single Chain Fv ◽  
Single Chain Fv Fragment

scholarly journals The in vivo aggregation of single chain Fv fragment can be replicated in vitro

2003 ◽  
Vol 43 (supplement) ◽  
pp. S62
Author(s):  
M. Umetsu ◽  
K. Tsumoto ◽  
Kumar Ashish ◽  
S. Nitta ◽  
Y. Tanaka ◽  
...  
Keyword(s):  
Single Chain ◽  
Single Chain Fv ◽  
Single Chain Fv Fragment

scholarly journals Gallium-68-Labeled Anti-HER2 Single-Chain Fv Fragment: Development and In Vivo Monitoring of HER2 Expression

2014 ◽  
Vol 17 (1) ◽  
pp. 102-110 ◽  
Author(s):  
Masashi Ueda ◽  
Hayato Hisada ◽  
Takashi Temma ◽  
Yoichi Shimizu ◽  
Hiroyuki Kimura ◽  
...  
Keyword(s):  
Her2 Expression ◽  
Single Chain ◽  
In Vivo Monitoring ◽  
Single Chain Fv ◽  
Gallium 68 ◽  
Single Chain Fv Fragment

Toward in Vivo Imaging of Heart Disease Using a Radiolabeled Single-Chain Fv Fragment Targeting Tenascin-C

2011 ◽  
Vol 83 (23) ◽  
pp. 9123-9130 ◽  
Author(s):  
Norihiro Kobayashi ◽  
Kenichi Odaka ◽  
Tomoya Uehara ◽  
Kyoko Imanaka-Yoshida ◽  
Yoshinori Kato ◽  
...  
Keyword(s):  
Heart Disease ◽  
In Vivo Imaging ◽  
Single Chain ◽  
Tenascin C ◽  
Single Chain Fv ◽  
Single Chain Fv Fragment

Anti-Human Epidermal Growth Factor Receptor 2 Single-Chain Fv Fragment-Decorated DM1 Nanoparticles for Specific Targeting of Human Epidermal Growth Factor Receptor 2-Positive Breast Tumor Cells

2021 ◽  
Vol 17 (3) ◽  
pp. 447-455
Author(s):  
Ling Ni ◽  
You-Xin Li
Keyword(s):  
Breast Cancer ◽  
Epidermal Growth Factor Receptor ◽  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Growth Factor Receptor ◽  
Single Chain ◽  
Human Epidermal Growth Factor ◽  
Single Chain Fv ◽  
Epidermal Growth ◽  
Single Chain Fv Fragment

Purpose: Although monoclonal antibodies are used to decorate nanoparticles to target specific cells, penetration of tumor tissues by monoclonal antibodies is limited by their large size. Therefore, we prepared DM1 nanoparticles decorated with the small anti-HER2 single-chain Fv fragment (scFvHER2) of trastuzumab (TMAB) for targeting to human epidermal growth factor receptor 2 (HER2) overexpressing in breast cancer effectively. Methods: ScFvHER2 fragment was coupled with DM1 nanoparticles (NPs) via covalent thiol-maleimide linkages. Their physicochemical properties, uptake by cells, and toxicity to tumor cells were investigated. Their vivo biodistribution was assessed employing liquid chromatographytandem mass spectrometry, while their antitumor activity was investigated in nude mice burdened with BT-474 tumor. Results: Viability of BT-474 cells incubated with scFvHER2-DM1-Nanoparticles (scFv-DM1-NPs) was significantly lower than that of BT-474 cell treated with TMAB-DM1-Nanoparticles (TMAB-DM1-NPs) (P < 0 05). Uptake by cells of scFvDM1-NPs was significantly higher than TMAB-DM1-NPs (P < 0 01). Accumulation of scFv-DM1-NPs in tumor tissue was notably higher than TMAB-DM1-NPs (P < 0 05). scFv-DM1-NPs exhibited improved antitumor effects compared to TMABDM1-NPs (P < 0 05), showing a tumor inhibition rate of more than 70%. Conclusions: ScFvHER2 fragment could serve as a more effective targeting ligand than TMAB, and scFv-DM1-NPs could be developed as a possible drug delivery system to target HER2-positive breast cancer.

scholarly journals Engineered Fragments of the PSMA-Specific 5D3 Antibody and Their Functional Characterization

2020 ◽  
Vol 21 (18) ◽  
pp. 6672
Author(s):  
Zora Novakova ◽  
Nikola Belousova ◽  
Catherine A. Foss ◽  
Barbora Havlinova ◽  
Marketa Gresova ◽  
...  
Keyword(s):  
Functional Characterization ◽  
Xenograft Model ◽  
In Vitro Assays ◽  
Experimental Therapy ◽  
Single Chain ◽  
Single Chain Fv ◽  
Murine Xenograft Model ◽  
Single Target

Prostate-Specific Membrane Antigen (PSMA) is an established biomarker for the imaging and experimental therapy of prostate cancer (PCa), as it is strongly upregulated in high-grade primary, androgen-independent, and metastatic lesions. Here, we report on the development and functional characterization of recombinant single-chain Fv (scFv) and Fab fragments derived from the 5D3 PSMA-specific monoclonal antibody (mAb). These fragments were engineered, heterologously expressed in insect S2 cells, and purified to homogeneity with yields up to 20 mg/L. In vitro assays including ELISA, immunofluorescence and flow cytometry, revealed that the fragments retain the nanomolar affinity and single target specificity of the parent 5D3 antibody. Importantly, using a murine xenograft model of PCa, we verified the suitability of fluorescently labeled fragments for in vivo imaging of PSMA-positive tumors and compared their pharmacokinetics and tissue distribution to the parent mAb. Collectively, our data provide an experimental basis for the further development of 5D3 recombinant fragments for future clinical use.

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