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Predicate | Object |
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
3-4
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pubmed:dateCreated |
1981-3-27
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pubmed:abstractText |
Previous studies on the determination of the complete amino acid sequence of H-2 molecules have relied on alloantisera to purify the H-2 molecules by immunoprecipitation. In this study, directed toward the determination of the complete amino acid sequence of the H-2Db molecule, it was found that all H-2.2 alloantisera examined had very low antibody titers when analyzed by immunoprecipitation and that most of these antisera reacted with not only the H-2Db heavy chain but also with a 75,000 dalton virus-associated glycoprotein (gp 75). The problems encountered with conventional alloantisera were surmounted by utilizing a monoclonal antibody from hybridoma B22-249R1 obtained as described by Hämmerling et al. (1979) and maintained in ascites form in (CBA x Balb/c) F1 mice. This antibody reacted with only 45,000 molecular weight material and had the added advantage of requiring only 0.1 ml of hybridoma ascites fluid to bind all of the H-2Db antigen from 3 x 10(8) EK-4 tumor cells. Biochemical examination of the purified H-2Db molecule showed that it possessed overall structural properties similar to other previously characterized H-2 molecules. An NH2-terminal amino acid sequence was determined for the H-2Db molecule for 28 residues and this sequence was compared to the complete NH2-terminal sequences of the H-2Kb and H-2Dd molecules, and the partial NH2-terminal sequence for the H-2Ld molecule; these comparisons indicated 82%, 75% and 100% homology respectively. In addition, these preliminary studies indicated that there was little or no beta-2-microglobulin (beta 2 mu) in the immunoprecipitates with the H2-2Db heavy chain. This lack of beta 2 mu in the immunoprecipitates was found to be true for alloantisera directed against private or public specificities, a xenoantiserum (sheep anti-H-2b), or hybridoma antibody used in the immunoprecipitation.
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pubmed:language |
eng
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pubmed:journal | |
pubmed:citationSubset |
IM
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pubmed:chemical | |
pubmed:status |
MEDLINE
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pubmed:issn |
0022-1759
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pubmed:author | |
pubmed:issnType |
Print
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pubmed:volume |
37
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
287-99
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pubmed:dateRevised |
2006-11-15
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pubmed:meshHeading |
pubmed-meshheading:7005347-Amino Acid Sequence,
pubmed-meshheading:7005347-Animals,
pubmed-meshheading:7005347-Antibodies, Neoplasm,
pubmed-meshheading:7005347-Antigen-Antibody Reactions,
pubmed-meshheading:7005347-Clone Cells,
pubmed-meshheading:7005347-H-2 Antigens,
pubmed-meshheading:7005347-Hybrid Cells,
pubmed-meshheading:7005347-Immunoglobulin Allotypes,
pubmed-meshheading:7005347-Mice,
pubmed-meshheading:7005347-Mice, Inbred Strains,
pubmed-meshheading:7005347-Molecular Weight,
pubmed-meshheading:7005347-Neoplasms, Experimental,
pubmed-meshheading:7005347-beta 2-Microglobulin
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pubmed:year |
1980
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pubmed:articleTitle |
Comparison of alloantisera and hybridoma antibody for purification of the H-2Db murine histocompatibility antigen and preliminary molecular characterization of this antigen.
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pubmed:publicationType |
Journal Article,
Comparative Study
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