Production of specific antibodies against protein A fusion proteins

The gene for Staphylococcal protein A was fused to the coding sequence of bacterial beta‐galactosidase, alkaline phosphatase and human insulin‐like growth factor I (IGF‐I). The fusion proteins, expressed in bacteria, were purified by affinity chromatography on IgG‐Sepharose and antibodies were raise...

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Bibliographic Details
Published inThe EMBO journal Vol. 5; no. 9; pp. 2393 - 2398
Main Authors Löwenadler, B., Nilsson, B., Abrahmsén, L., Moks, T., Ljungqvist, L., Holmgren, E., Paleus, S., Josephson, S., Philipson, L., Uhlén, M.
Format Journal Article
LanguageEnglish
Published London Nature Publishing Group 01.09.1986
Subjects
Alkaline Phosphatase - analysis
Amino Acid Sequence
Antibodies
Antigen-Antibody Complex
Bacteriology
Base Sequence
beta-Galactosidase - analysis
Biological and medical sciences
Escherichia coli - genetics
Fundamental and applied biological sciences. Psychology
Genes
Humans
Insulin-Like Growth Factor I - analysis
Microbiology
Pathogenicity, virulence, toxins, bacteriocins, pyrogens, host-bacteria relations, miscellaneous strains
Recombinant Fusion Proteins - analysis
Recombinant Fusion Proteins - genetics
Recombinant Fusion Proteins - immunology
Recombinant Proteins - analysis
Staphylococcal Protein A - analysis
Staphylococcal Protein A - immunology
Proteins
Antibody
Production
Molecular hybrid
Bacteria
Micrococcales
Micrococcaceae
Staphylococcus
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Summary:The gene for Staphylococcal protein A was fused to the coding sequence of bacterial beta‐galactosidase, alkaline phosphatase and human insulin‐like growth factor I (IGF‐I). The fusion proteins, expressed in bacteria, were purified by affinity chromatography on IgG‐Sepharose and antibodies were raised in rabbits. All three fusion proteins elicited specific antibodies against both the inserted protein sequences and the protein A moiety. In the case of IGF‐I, the protein A moiety in the fusion protein may act as an adjuvant since native IGF‐I alone is a poor immunogen. The results suggest that the protein A fusion system can be used for efficient antibody production against peptides or proteins expressed from cloned or synthetic genes. To facilitate such gene fusions a set of optimized vectors have been constructed.
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SourceType-Scholarly Journals-1
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ISSN:0261-4189
1460-2075
DOI:10.1002/j.1460-2075.1986.tb04509.x