• Login
    View Item 
    •   Home
    • The Manchester Institute Cancer Research UK
    • All Paterson Institute for Cancer Research
    • View Item
    •   Home
    • The Manchester Institute Cancer Research UK
    • All Paterson Institute for Cancer Research
    • View Item
    JavaScript is disabled for your browser. Some features of this site may not work without it.

    Browse

    All of ChristieCommunitiesTitleAuthorsIssue DateSubmit DateSubjectsThis CollectionTitleAuthorsIssue DateSubmit DateSubjects

    My Account

    LoginRegister

    Local Links

    The Christie WebsiteChristie Library and Knowledge Service

    Statistics

    Display statistics

    Isolation of a high affinity scFv from a monoclonal antibody recognising the oncofoetal antigen 5T4.

    • CSV
    • RefMan
    • EndNote
    • BibTex
    • RefWorks
    Authors
    Shaw, David M
    Embleton, Jim
    Westwater, C
    Ryan, Matthew G
    Myers, Kevin A
    Kingsman, Susan M
    Carroll, M W
    Stern, Peter L
    Affiliation
    CRC Immunology Group, Paterson Institute for Cancer Research, Christie Hospital, Manchester, UK.
    Issue Date
    2000-12-15
    
    Metadata
    Show full item record
    Abstract
    The oncofoetal antigen 5T4 is a 72 kDa glycoprotein expressed at the cell surface. It is defined by a monoclonal antibody, mAb5T4, that recognises a conformational extracellular epitope in the molecule. Overexpression of 5T4 antigen by tumours of several types has been linked with disease progression and poor clinical outcome. Its restricted expression in non-malignant tissue makes 5T4 antigen a suitable target for the development of antibody directed therapies. The use of murine monoclonal antibodies for targeted therapy allows the tumour specific delivery of therapeutic agents. However, their use has several drawbacks, including a strong human anti-mouse immune (HAMA) response and limited tumour penetration due to the size of the molecules. The use of antibody fragments leads to improved targeting, pharmacokinetics and a reduced HAMA. A single chain antibody (scFv) comprising the variable regions of the mAb5T4 heavy and light chains has been expressed in Escherichia coli. The addition of a eukaryotic leader sequence allowed production in mammalian cells. The two 5T4 single chain antibodies, scFv5T4WT19 and LscFv5T4, described the same pattern of 5T4 antigen expression as mAb5T4 in normal human placenta and by FACS. Construction of a 5T4 extracellular domain-IgGFc fusion protein and its expression in COS-7 cells allowed the relative affinities of the antibodies to be compared by ELISA and measured in real time using a biosensor based assay. MAb5T4 has a high affinity, K(D)=1.8x10(-11) M, as did both single chain antibodies, scFv5T4WT19 K(D)=2.3x10(-9) M and LscFv5T4 K(D)=7.9x10(-10) M. The small size of this 5T4 specific scFv should allow construction of fusion proteins with a range of biological response modifiers to be prepared whilst retaining the improved pharmacokinetic properties of scFvs.
    Citation
    Isolation of a high affinity scFv from a monoclonal antibody recognising the oncofoetal antigen 5T4. 2000, 1524 (2-3):238-46 Biochim. Biophys. Acta
    Journal
    Biochimica et Biophysica Acta
    URI
    http://hdl.handle.net/10541/86122
    PubMed ID
    11113573
    Type
    Article
    Language
    en
    ISSN
    0006-3002
    Collections
    All Paterson Institute for Cancer Research

    entitlement

    Related articles

    • Characterization of the murine 5T4 oncofoetal antigen: a target for immunotherapy in cancer.
    • Authors: Woods AM, Wang WW, Shaw DM, Ward CM, Carroll MW, Rees BR, Stern PL
    • Issue date: 2002 Aug 15
    • Pharmacokinetics and biodistribution of genetically-engineered antibodies.
    • Authors: Colcher D, Pavlinkova G, Beresford G, Booth BJ, Choudhury A, Batra SK
    • Issue date: 1998 Dec
    • High-affinity recombinant phage antibodies to the pan-carcinoma marker epithelial glycoprotein-2 for tumour targeting.
    • Authors: Roovers RC, Henderikx P, Helfrich W, van der Linden E, Reurs A, de Bruïne AP, Arends JW, de Leij L, Hoogenboom HR
    • Issue date: 1998 Dec
    • Guided selection of a pan carcinoma specific antibody reveals similar binding characteristics yet structural divergence between the original murine antibody and its human equivalent.
    • Authors: Beiboer SH, Reurs A, Roovers RC, Arends JW, Whitelegg NR, Rees AR, Hoogenboom HR
    • Issue date: 2000 Feb 25
    • Anti-estradiol-17beta single-chain Fv fragments: Generation, characterization, gene randomization, and optimized phage display.
    • Authors: Kobayashi N, Kato Y, Oyama H, Taga S, Niwa T, Sun P, Ohtoyo M, Goto J
    • Issue date: 2008 Dec 22
    DSpace software (copyright © 2002 - 2023)  DuraSpace
    Quick Guide | Contact Us
    Open Repository is a service operated by 
    Atmire NV
     

    Export search results

    The export option will allow you to export the current search results of the entered query to a file. Different formats are available for download. To export the items, click on the button corresponding with the preferred download format.

    By default, clicking on the export buttons will result in a download of the allowed maximum amount of items.

    To select a subset of the search results, click "Selective Export" button and make a selection of the items you want to export. The amount of items that can be exported at once is similarly restricted as the full export.

    After making a selection, click one of the export format buttons. The amount of items that will be exported is indicated in the bubble next to export format.