Linked Life Data

11113573

Source:http://linkedlifedata.com/resource/pubmed/id/11113573

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
2-3
pubmed:dateCreated
2001-1-16
pubmed:abstractText
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.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Dec
pubmed:issn
0006-3002
pubmed:author
pubmed:issnType
Print
pubmed:day
15
pubmed:volume
1524
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
238-46
pubmed:dateRevised
2006-11-15
pubmed:meshHeading
pubmed-meshheading:11113573-Amino Acid Sequence, pubmed-meshheading:11113573-Antibodies, Monoclonal, pubmed-meshheading:11113573-Antigens, Neoplasm, pubmed-meshheading:11113573-Base Sequence, pubmed-meshheading:11113573-Cloning, Molecular, pubmed-meshheading:11113573-Enzyme-Linked Immunosorbent Assay, pubmed-meshheading:11113573-Female, pubmed-meshheading:11113573-Gene Therapy, pubmed-meshheading:11113573-Humans, pubmed-meshheading:11113573-Immunoglobulin Fragments, pubmed-meshheading:11113573-Immunoglobulin Variable Region, pubmed-meshheading:11113573-Immunohistochemistry, pubmed-meshheading:11113573-Membrane Glycoproteins, pubmed-meshheading:11113573-Molecular Sequence Data, pubmed-meshheading:11113573-Mutation, pubmed-meshheading:11113573-Placenta, pubmed-meshheading:11113573-Recombinant Fusion Proteins, pubmed-meshheading:11113573-Stomach Neoplasms, pubmed-meshheading:11113573-Surface Plasmon Resonance
pubmed:year
2000
pubmed:articleTitle
Isolation of a high affinity scFv from a monoclonal antibody recognising the oncofoetal antigen 5T4.
pubmed:affiliation
CRC Immunology Group, Paterson Institute for Cancer Research, Christie Hospital, Manchester, UK.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't