Expression of Recombinant Mouse/Human Chimeric Antibody Specific to Human GMP-140/P-selectin

Kurome, Toru; Katayama, Masahiko; Murakami, Keiko; Hashino, Kimikazu; Kamihagi, K; Yasumoto, Masazumi; Kato, Ikunoshin

doi:10.1093/oxfordjournals.jbchem.a124383

Cited by 3 publications

(2 citation statements)

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“…However, very few recombinant antibodies are available to study the clinical function of P-selectin except for therapeutic attempts in the form of monoclonal antibodies and chimeric antibodies. 10,15,19,32,34 Presently, there have not been any studies using P-selectin as an imaging target in cancer, partly due to the lack of specific desired antibodies to the protein. For these reasons, the aim of this study is to develop antihuman P-selectin scFvs that can be used for radiation-guided tumor imaging.…”

Section: Therapeutic Antibodiesmentioning

confidence: 99%

Radiation-Guided P-Selectin Antibody Targeted to Lung Cancer

Hariri

Zhang

et al. 2008

Ann Biomed Eng

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Purpose-P-selectin expression is significantly increased in tumor microvasculature following exposure to ionizing radiation. The purpose of this study was to image radiation-induced P-selectin expression in vivo using optical imaging and gamma camera imaging in a heterotopic lung cancer model by using ScFv antibodies to P-selectin.Procedures-In vitro studies using endothelial cells were done using 3 Gy radiation and selected ScFv antibodies to P-selectin. In vivo studies were performed using Lewis lung carcinoma cells subcutaneously injected into the hind limbs of nude mice. Mice were treated with 6 Gy radiation and sham radiation 10 days post-inoculation. P-selectin expression was assessed with near-infrared imaging using Cy7 labeled antibody, and gamma camera imaging using 111 In-DTPA labeled antibody.Results-In vitro studies showed antibody binding to P-selectin in radiation treated endothelial cells. In vivo optical imaging and gamma camera imaging studies showed significant tumor-specific binding to P-selectin in irradiated tumors compared to unirradiated tumors.Conclusions-Optical imaging and gamma camera imaging are effective methods for visualizing in vivo targeting of radiation-induced P-selectin in lung tumors. This study suggests that fluorescentlabeled and radiolabeled ScFv antibodies can be used to target radiation-induced P-selectin for the tumor-specific delivery of therapeutic drugs and radionuclides in vivo.

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Section: Therapeutic Antibodiesmentioning

confidence: 99%

Radiation-Guided P-Selectin Antibody Targeted to Lung Cancer

Hariri

Zhang

et al. 2008

Ann Biomed Eng

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“…Thrombi, especially arterial thrombi, contain a large number of activated platelets. It is expected that an antibody that reacts specifically with P-selectin would be directed selectively to these activated platelets in thrombi (3,4).…”

Section: Introductionmentioning

confidence: 99%

Construction and Expression of Mouse-human Chimeric Antibody SZ-51 Specific for Activated Platelet P-selectin

Gu¹,

Liu

Xia

et al. 1997

Thromb Haemost

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SummaryA murine monoclonal (mAb) SZ-51 specific for human P-selectin may be used for in vivo thrombus imaging and for the targeting of fibrinolytic agents to thrombi. In order to reduce the immunogenicity of the murine mAb SZ-51 in humans, we cloned and sequenced the cDNAs encoding the variable region of mAb SZ-51 in order to develop mouse/human chimeric reagents. The E. coli expression vector. pHENl-SZ51 Fab/Hu was constructed by fusing the variable regions of mAb SZ-51 with human IgG γICHI and Cκ genes. The constructs were introduced into E. coli HB2151 for expression of soluble chimeric Fab fragment. We also constructed two fusion products by joining the variable regions of mouse antibody to the appropriate constant regions of human Igγl and κ. These chimeras were cloned into two eukaryotic selectable expression vectors separately, which were then cotransfected into a non-Ig secreting murine myeloma line SP2/0 with lipofectin reagent. Six cell lines remained positive for Ig secretion. The highest producing cell line, which showed stable integration and expression at 5 mg/1 of culture, was selected for the large scale production of chimeric antibody. Immunoblotting analysis demonstrated that both of the chimeric antibodies (SZ51Fab/Hu, SZ51/Hu) in the culture supernatants, like the native mAb SZ-51, bind P-selectin. In addition, the whole chimeric antibody can compete for binding to activated platelets with murine SZ-51. Therefore, the SZ-51 chimeric antibody may be a potential agent for diagnosis and treatment of thrombotic diseases in the future.

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Design of Human-type monoclonal antibody and Its Recent Clinical Application

Katayama

1996

Jpn J Thromb Hemost

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Expression of Recombinant Mouse/Human Chimeric Antibody Specific to Human GMP-140/P-selectin

Cited by 3 publications

References 0 publications

Radiation-Guided P-Selectin Antibody Targeted to Lung Cancer

Radiation-Guided P-Selectin Antibody Targeted to Lung Cancer

Construction and Expression of Mouse-human Chimeric Antibody SZ-51 Specific for Activated Platelet P-selectin

Design of Human-type monoclonal antibody and Its Recent Clinical Application

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