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Predicate | Object |
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
1
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pubmed:dateCreated |
1991-5-23
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pubmed:abstractText |
The binding of several oligosaccharide haptens by a monoclonal antibody, Se155-4, specific for Salmonella serogroup B O-antigen was studied by titration microcalorimetry. In the software developed by Wiseman et al. [Wiseman, T., Williston, S. & Brandts, J.F. (1989) Anal. Biochem. 17, 131-137] the number of binding sites/macromolecule is one of the optional regression parameters in the non-linear least-squares analysis of the calorimetric data. Instead, an approach was adopted in which the concentration of binding sites was treated as a regression parameter, obviating the requirement for precise values of antibody absorption coefficients and minimizing effects due to partially inactive antibody preparations. Furthermore, performing the least-squares analysis in two steps, first using a differential heat mode and then an integral heat mode, was shown to yield the most accurate results. The technique gave accurate results using not more than 1-2 mumol ligand and less than 7 mg antibody. Haptens 2-5 were oligomers of the O-antigenic repeating unit varying in chain length by 2-5 repeating units and a trisaccharide glycoside 1, which filled the binding site. The latter hapten exhibited a favourable entropy contribution to binding (delta Go = -31 kJ.mol-1; delta Ho = -21 kJ.mol-1 and -T delta So -10 kJ.mol-1), while all four oligomers 2-5 showed a constant binding energy delta Go = -33 kJ.mol-1, composed of increasingly stronger enthalpy forces compensated by an increasingly unfavourable entropy contribution. These observations are compared with results from enzyme immunoassays and a high-resolution crystal structure for the dodecasaccharide 3 bound to the Fab derived from Se155-4.
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pubmed:language |
eng
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pubmed:journal | |
pubmed:citationSubset |
IM
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pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/Antibodies, Monoclonal,
http://linkedlifedata.com/resource/pubmed/chemical/Antigens, Bacterial,
http://linkedlifedata.com/resource/pubmed/chemical/Epitopes,
http://linkedlifedata.com/resource/pubmed/chemical/Ligands,
http://linkedlifedata.com/resource/pubmed/chemical/Oligosaccharides,
http://linkedlifedata.com/resource/pubmed/chemical/Polysaccharides, Bacterial
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pubmed:status |
MEDLINE
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pubmed:month |
Apr
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pubmed:issn |
0014-2956
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pubmed:author | |
pubmed:issnType |
Print
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pubmed:day |
10
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pubmed:volume |
197
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
239-46
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pubmed:dateRevised |
2011-11-17
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pubmed:meshHeading |
pubmed-meshheading:1707812-Antibodies, Monoclonal,
pubmed-meshheading:1707812-Antigens, Bacterial,
pubmed-meshheading:1707812-Binding Sites, Antibody,
pubmed-meshheading:1707812-Calorimetry,
pubmed-meshheading:1707812-Carbohydrate Conformation,
pubmed-meshheading:1707812-Carbohydrate Sequence,
pubmed-meshheading:1707812-Enzyme-Linked Immunosorbent Assay,
pubmed-meshheading:1707812-Epitopes,
pubmed-meshheading:1707812-Ligands,
pubmed-meshheading:1707812-Models, Molecular,
pubmed-meshheading:1707812-Molecular Sequence Data,
pubmed-meshheading:1707812-Oligosaccharides,
pubmed-meshheading:1707812-Polysaccharides, Bacterial,
pubmed-meshheading:1707812-Regression Analysis,
pubmed-meshheading:1707812-Salmonella
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pubmed:year |
1991
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pubmed:articleTitle |
Sensitive titration microcalorimetric study of the binding of Salmonella O-antigenic oligosaccharides by a monoclonal antibody.
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pubmed:affiliation |
Institute for Biological Sciences, National Research Council of Canada, Ottawa.
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pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
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