scholarly journals Clinical Applications of Phage-Derived sFvs and sFv Fusion Proteins

2000 ◽  
Vol 16 (1-2) ◽  
pp. 53-62 ◽  
Author(s):  
K. A. Chester ◽  
J. Bhatia ◽  
G. Boxer ◽  
S. P. Cooke ◽  
A. A. Flynn ◽  
...  
Keyword(s):  
Fusion Proteins ◽  
Gamma Camera ◽  
Single Chain ◽  
Enzyme Prodrug Therapy ◽  
Radioimmunoguided Surgery ◽  
Patients With Cancer ◽  
Carboxypeptidase G2 ◽  
Single Chain Fv ◽  
Surgery Trial ◽  
Colorectal Adenocarcinomas

Single chain Fv antibodies (sFvs) have been produced from filamentous bacteriophage libraries obtained from immunised mice. MFE-23, the most characterised of these sFvs, is reactive with carcinoembryonic antigen (CEA), a glycoprotein that is highly expressed in colorectal adenocarcinomas. MFE-23 has been expressed in bacteria and purified in our laboratory for two clinical trials; a gamma camera imaging trial using123I-MFE-23 and a radioimmunoguided surgery trial using125I-MFE-23, where tumour deposits are detected by a hand-held probe during surgery. Both these trials show MFE-23 is safe and effective in localising tumour deposits in patients with cancer. We are now developing fusion proteins which use MFE-23 to deliver a therapeutic moiety; MFE-23::CPG2 targets the enzyme carboxypeptidase G2 (CPG2) for use in the ADEPT (antibody directed enzyme prodrug therapy) system and MFE::TNFα aims to reduce sequestration and increase tumor concentrations of systemically administered TNFα.

A vaccine approach for breast cancer immunotherapy by targeting HER2/neu antigen to B cells via CD19 molecule.

2011 ◽  
Vol 29 (27_suppl) ◽  
pp. 287-287
Author(s):  
D. Xiang ◽  
Y. Ma ◽  
C. Ding ◽  
G. H. Kloecker ◽  
J. Yan
Keyword(s):  
Breast Cancer ◽  
B Cells ◽  
Fusion Protein ◽  
Tumor Cells ◽  
Fusion Proteins ◽  
In Vivo Studies ◽  
Single Chain ◽  
Tumor Vaccines ◽  
Single Chain Fv ◽  
Her 2

287 Background: Trastuzumab has been proven to be effective in the immunotherapy for Her-2/neu-positive breast cancer. However, this adoptively transferred therapeutic Ab must be continuously given to the patient with huge financial cost. Thus, it would be desirable if tumor vaccines could elicit long-lasting trastuzumab-like Ab. Methods: Constructing fusion protein of anti-mouse CD19 single chain variable fragment (anti-CD19-scFv) with human Her-2/neu extracellular domain P3-4 (P3-4) by extracting total RNA from Rat anti-mouse CD19 hybridoma (1D3) and first strand cDNA was synthesized accordingly. VH and VL were amplified using designed primers. Single chain Fv (VL-VH) was then synthesized by overlapping PCR. Then constructs were sequenced and cloned into pET-20b(+) vectors. Fragments P3-4 of extracellular domains of human Her-2/neu was cloned from pcDNA3.1-Her2 and then cloned into pET-20b(+)-scFv. Series of constructs were expressed and purified according to standard protocol and verified by Western Blot. Results: Both in vitro and in vivo studies demonstrated that fusion proteins anti-CD19-scFv and anti-CD19-scFv-P3-4 but not P3-4 could specifically bind to B cells with high affinity. Mice immunized with anti-CD19-scFv-P3-4 secreted higher titers of IgG and IgM than those from controls (p<0.05). Studies also demonstrated that sera from anti-CD19-scFv-P3-4 but not anti-CD19-scFv or P3-4 immunized mice stained with Her-2/neu expressing SKOV-3 tumor cells. These Abs also competitively inhibited trastuzumab-mediated Ag binding, suggesting that trastuzumab-like Ab responses were elicited. WT mice immunized with anti-CD19-scFv-P3-4 fusion protein then challenged with D2/F2-Her-2 mammary tumor cells showed significantly reduced tumor burden compared to those immunized with control fusion proteins (p<0.05) and had enhanced median overall survival to 45 days versus 34 days in WT mice immunized with either anti-CD19-scFv or P3-4. Conclusions: Targeting of Her-2/neu antigens to B cells stimulates Th-dependent humoral immune responses with anti tumor effect in mouse model. These findings provide a novel avenue for successful development of breast cancer vaccination strategy and help to fight for cancer.

Single-Chain Fv Fusion Proteins Suitable as Coating and Detecting Reagents in a Double Antibody Sandwich Enzyme-Linked Immunosorbent Assay

1997 ◽  
Vol 249 (2) ◽  
pp. 219-227 ◽  
Author(s):  
Randolf J. Kerschbaumer ◽  
Sonja Hirschl ◽  
Andrea Kaufmann ◽  
Martin Ibl ◽  
Renate Koenig ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Fusion Proteins ◽  
Enzyme Linked Immunosorbent Assay ◽  
Single Chain ◽  
Double Antibody ◽  
Single Chain Fv

Fluobodies: green fluorescent single-chain Fv fusion proteins

1999 ◽  
Vol 230 (1-2) ◽  
pp. 121-130 ◽  
Author(s):  
Remko A Griep ◽  
Charlotte van Twisk ◽  
Jan M van der Wolf ◽  
Arjen Schots
Keyword(s):  
Fusion Proteins ◽  
Single Chain ◽  
Single Chain Fv ◽  
Green Fluorescent

An ELISA for the detection of TIMP-1 based on recombinant single chain Fv fusion proteins

2003 ◽  
Vol 335 (1-2) ◽  
pp. 49-57 ◽  
Author(s):  
Gordana Wozniak ◽  
Eva Obermayr ◽  
Matjaz Jeras ◽  
Mio Knezevic ◽  
Florian Rüker
Keyword(s):  
Fusion Proteins ◽  
Single Chain ◽  
Single Chain Fv

scholarly journals Antibody-based delivery of interleukin-9 to neovascular structures: Therapeutic evaluation in cancer and arthritis

2021 ◽  
pp. 153537022098157
Author(s):  
Baptiste Gouyou ◽  
Tiziano Ongaro ◽  
Samuele Cazzamalli ◽  
Roberto De Luca ◽  
Anne Kerschenmeyer ◽  
...  
Keyword(s):  
Targeted Delivery ◽  
Fusion Proteins ◽  
Therapeutic Benefit ◽  
Negative Control ◽  
Lymphoid Cells ◽  
Single Chain ◽  
Single Chain Fv ◽  
Disease Site ◽  
Group 2 ◽  
Interleukin 9

Interleukin-9 is a cytokine with multiple functions, including the ability to activate group 2 innate lymphoid cells, which has been postulated to be therapeutically active in mouse models of arthritis. Similarly, interleukin-9 has been suggested to play an important role in tumor immunity. Here, we describe the cloning, expression, and characterization of three fusion proteins based on murine interleukin-9 and the F8 antibody, specific to the alternatively spliced EDA domain of fibronectin. EDA is strongly expressed in cancer and in various arthritic conditions, while being undetectable in the majority of healthy organs. Interleukin-9-based fusion proteins with an irrelevant antibody specific to hen egg lysozyme served as negative control in our study. The fusion proteins were characterized by quantitative biodistribution analysis in tumor-bearing mice using radioiodinated protein preparations. The highest tumor uptake and best tumor:organ ratios were observed for a format, in which the interleukin-9 moiety was flanked by two units of the F8 antibody in single-chain Fv format. Biological activity of interleukin-9 was retained when the payload was fused to antibodies. However, the targeted delivery of interleukin-9 to the disease site resulted in a modest anti-tumor activity in three different murine models of cancer (K1735M2, CT26, and F9), while no therapeutic benefit was observed in a collagen induced model of arthritis. Collectively, these results confirm the possibility to deliver interleukin-9 to the site of disease but cast doubts about the alleged therapeutic activity of this cytokine in cancer and arthritis, which has been postulated in previous publications.

scholarly journals Antibody-based delivery of Interleukin-9 to neovascular structures: therapeutic evaluation in cancer and arthritis

2020 ◽  
Author(s):  
Baptiste Gouyou ◽  
Tiziano Ongaro ◽  
Samuele Cazzamalli ◽  
Roberto De Luca ◽  
Anne Kerschenmeyer ◽  
...  
Keyword(s):  
Targeted Delivery ◽  
Fusion Proteins ◽  
Therapeutic Benefit ◽  
Negative Control ◽  
Lymphoid Cells ◽  
Single Chain ◽  
Single Chain Fv ◽  
Disease Site ◽  
Group 2 ◽  
Interleukin 9

ABSTRACTInterleukin-9 (IL9) is a cytokine with multiple functions, including the ability to activate group 2 innate lymphoid cells (ILC2s), which has been postulated to be therapeutically active in mouse models of arthritis. Similarly, IL9 has been suggested to play an important role in tumor immunity. Here, we describe the cloning, expression and characterization of three fusion proteins based on murine IL9 and the F8 antibody, specific to the alternatively-spliced EDA domain of fibronectin. EDA is strongly expressed in cancer and in various arthritic conditions, while being undetectable in the majority of healthy organs. IL9-based fusion proteins with an irrelevant antibody specific to hen egg lysozyme served as negative control in our study. The fusion proteins were characterized by quantitative biodistribution analysis in tumor-bearing mice using radioiodinated protein preparations. The highest tumor uptake and best tumor:organ ratios were observed for a format, in which the IL9 moiety was flanked by two units of the F8 antibody in single-chain Fv format. Biological activity of IL9 was retained when the payload was fused to antibodies. However, the targeted delivery of IL9 to the disease site resulted in a modest anti-tumor activity in three different murine models of cancer (K1735M2, CT26 and F9), while no therapeutic benefit was observed in a collagen induced model of arthritis. Collectively, these results confirm the possibility to deliver IL9 to the site of disease but cast doubts about the alleged therapeutic activity of this cytokine in cancer and arthritis, which has been postulated in previous publications.

Sign in / Sign up

Export Citation Format

Share Document