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rdf:type |
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lifeskim:mentions |
umls-concept:C0003241,
umls-concept:C0015272,
umls-concept:C0033268,
umls-concept:C0037628,
umls-concept:C0162801,
umls-concept:C0231449,
umls-concept:C0521009,
umls-concept:C1514562,
umls-concept:C1880389,
umls-concept:C1883204,
umls-concept:C1883221,
umls-concept:C2700116
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pubmed:issue |
1
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pubmed:dateCreated |
1997-10-15
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pubmed:databankReference |
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pubmed:abstractText |
The cDNAs for the two variable domains of the antibody 9E10 were cloned from the hybridoma cell line. A chimeric 9E10 Fab fragment was produced in E. coli under control of the tightly controlled tetracycline promoter. The functional Fab fragment was isolated in a single step via a His6-tag, which also served for its recognition by a nickel chelate-alkaline phosphatase conjugate. Thus, the recombinant Fab fragment permitted the immunochemical detection of the myc tag in a sandwich ELISA. The dissociation constant for the interaction with the myc tag peptide was determined as 80 +/- 5 nM by fluorescence titration. In an attempt to produce the smaller 9E10 Fv fragment it was found that its V(H) domain alone can be readily isolated from E. coli as a soluble protein. This unusual behaviour may be explained by the 18 amino acid-long CDR-H3 and could be of value in the design of 'single domain' antibodies.
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pubmed:language |
eng
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pubmed:journal |
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pubmed:citationSubset |
IM
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pubmed:chemical |
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pubmed:status |
MEDLINE
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pubmed:month |
Sep
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pubmed:issn |
0014-5793
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:day |
1
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pubmed:volume |
414
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
33-8
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pubmed:dateRevised |
2005-11-17
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pubmed:meshHeading |
pubmed-meshheading:9305727-Amino Acid Sequence,
pubmed-meshheading:9305727-Animals,
pubmed-meshheading:9305727-Antibodies, Monoclonal,
pubmed-meshheading:9305727-Base Sequence,
pubmed-meshheading:9305727-Cloning, Molecular,
pubmed-meshheading:9305727-Enzyme-Linked Immunosorbent Assay,
pubmed-meshheading:9305727-Escherichia coli,
pubmed-meshheading:9305727-Genes, Immunoglobulin,
pubmed-meshheading:9305727-Histidine,
pubmed-meshheading:9305727-Hybridomas,
pubmed-meshheading:9305727-Immunoglobulin Fab Fragments,
pubmed-meshheading:9305727-Immunoglobulin Heavy Chains,
pubmed-meshheading:9305727-Immunoglobulin Variable Region,
pubmed-meshheading:9305727-Mice,
pubmed-meshheading:9305727-Molecular Sequence Data,
pubmed-meshheading:9305727-Mutagenesis, Site-Directed,
pubmed-meshheading:9305727-Peptides,
pubmed-meshheading:9305727-Polymerase Chain Reaction,
pubmed-meshheading:9305727-Proto-Oncogene Proteins c-myc,
pubmed-meshheading:9305727-Recombinant Fusion Proteins,
pubmed-meshheading:9305727-Sequence Analysis,
pubmed-meshheading:9305727-Solubility
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pubmed:year |
1997
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pubmed:articleTitle |
Sequence analysis and bacterial production of the anti-c-myc antibody 9E10: the V(H) domain has an extended CDR-H3 and exhibits unusual solubility.
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pubmed:affiliation |
Institut für Biochemie, Technische Hochschule, Darmstadt, Germany.
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pubmed:publicationType |
Journal Article
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