Back to Search
Start Over
A bifunctional fusion protein containing Fc-binding fragment B of staphylococcal protein A amino-terminal to antidigoxin single-chain Fv
- Source :
- Biochemistry. 29:8024-8030
- Publication Year :
- 1990
- Publisher :
- American Chemical Society (ACS), 1990.
-
Abstract
- A bifunctional molecule was genetically engineered which contained an amino-terminal effector domain that bound immunoglobulin Fc (fragment B of staphylococcal protein A) and a carboxyl-terminal domain that bound digoxin [a single-chain Fv (sFv)]. Effector and sFv binding properties were virtually identical with those of the parent molecules, despite the proximity of the FB to the sFv combining site. This finding is unprecedented since in all molecules of the natural immunoglobulin superfamily, the antigen binding domain is amino terminal to the effector domain. The FB-sFv sequence was encoded in a single synthetic gene and expressed as a 33,106 molecular weight protein in Escherichia coli. After purification, renaturation, and affinity isolation, yield of active fusion protein were 110 mg/L of fermented cells (18.5-g cell paste). Bifunctionality was confirmed by the ability of FB-sFv to cross-link IgG to digoxin-bovine serum albumin, as measured by plate assays and by Ouchterlony analysis. Analysis of the expressed fusion protein suggests that the sFv holds promise for the development of multifunctional, targetable single-chain proteins.
- Subjects :
- Digoxin
Protein Conformation
Recombinant Fusion Proteins
viruses
Immunoglobulin Fc
Molecular Sequence Data
Antibody Affinity
Protein Engineering
Biochemistry
Immunoglobulin Fab Fragments
Genes, Synthetic
Amino Acid Sequence
Staphylococcal Protein A
Gel electrophoresis
Base Sequence
biology
Effector
Nucleic acid sequence
Antibodies, Monoclonal
Fusion protein
Molecular biology
Peptide Fragments
Immunoglobulin Fc Fragments
biology.protein
Immunoglobulin superfamily
Binding Sites, Antibody
Antibody
Protein A
Protein Binding
Subjects
Details
- ISSN :
- 15204995 and 00062960
- Volume :
- 29
- Database :
- OpenAIRE
- Journal :
- Biochemistry
- Accession number :
- edsair.doi.dedup.....aefd9602434424299a5d647c0b51f989