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PMID: 2425364 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Genetic engineering of an H-2Dd/Q10b chimeric histocompatibility antigen: purification of soluble protein from transformant cell supernatants.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 83 ·No. 14 ·1986-07-00 ·Pages 5252-6

Margulies DH, Ramsey AL, Boyd LF, McCluskey J

Abstract

We have constructed a recombinant class I gene in which 5' sequences of H-2Dd are linked to the 3' half of a Qa subregion gene, Q10b. This hybrid gene would be expected to direct the synthesis of a protein containing the N and C1 domains of H-2Dd covalently linked to the C2 domain of the secreted, nonpolymorphic, Q10b antigen. Following DNA-mediated gene transfer into mouse L cells, transformants were analyzed by radiolabeling and immunoprecipitation. These cells secreted a molecule reactive with anti-H-2Dd monoclonal antibodies that identify epitopes on the N and C1 domains as well as with an anti-Q10 carboxyl-terminal peptide antiserum. The H-2Dd-derived antigen is associated with beta 2-microglobulin and is readily purified in milligram amounts from culture supernatants by immunoaffinity chromatography.

MeSH Terms
Animals Antibodies, Monoclonal/immunology Epitopes/immunology H-2 Antigens/genetics,immunology,isolation & purification Histocompatibility Antigen H-2D L Cells/analysis Mice Recombinant Proteins/genetics,isolation & purification Transformation, Genetic
Chemicals
Antibodies, Monoclonal Epitopes H-2 Antigens Histocompatibility Antigen H-2D Recombinant Proteins
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Margulies D H
Ramsey A L
Boyd L F
McCluskey J
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26 references, click to expand
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.83.14.5252
Published
1986-07-00
Pages
5252-6
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC323929
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.83.14.5252 PMC Full Text View journal details
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