| RRC ID |
33458
|
| Author |
Löwenadler B, Jansson B, Paleus S, Holmgren E, Nilsson B, Moks T, Palm G, Josephson S, Philipson L, Uhlén M.
|
| Title |
A gene fusion system for generating antibodies against short peptides.
|
| Journal |
Gene
|
| Abstract |
A novel method to obtain specific antibodies against short peptides is described, involving synthesis of the corresponding oligodeoxynucleotides followed by cloning into a new set of fusion vectors, pEZZ8 and pEZZ18, based on two synthetic IgG-binding domains (ZZ) of Staphylococcus aureus protein A. The soluble gene fusion product thus obtained, can be collected from the culture medium of Escherichia coli and rapidly recovered in a one-step procedure by IgG affinity chromatography. The system was used to express a fusion protein consisting of the two Z fragments and the C-terminal part [amino acids (aa) 57-70] of human insulin-like growth factor I (IGF-I). This 16-kDa protein was purified by affinity chromatography on IgG Sepharose and antibodies were raised in rabbits. The fusion protein elicited peptide-specific antibodies, as measured by solid-phase radioimmuno assay and Western blotting, reactive with both synthetic C-terminal peptide and the native human IGF-I protein. The results suggests that the gene fusion system can be used for efficient antibody production against short peptides encoded by synthetic oligodeoxynucleotides.
|
| Volume |
58(1)
|
| Pages |
87-97
|
| Published |
1987-1-1
|
| DOI |
10.1016/0378-1119(87)90032-1
|
| PII |
0378-1119(87)90032-1
|
| PMID |
3692175
|
| MeSH |
Amino Acid Sequence
Antibody Formation*
Base Sequence
Cloning, Molecular / methods*
Genetic Vectors
Humans
Insulin-Like Growth Factor I / genetics
Insulin-Like Growth Factor I / immunology
Molecular Sequence Data
Oligodeoxyribonucleotides / genetics
Peptides / genetics*
Peptides / immunology
Recombinant Fusion Proteins / genetics*
Recombinant Fusion Proteins / immunology
Recombinant Proteins / genetics*
|
| IF |
2.984
|
| Times Cited |
113
|
|
WOS Category
|
GENETICS & HEREDITY
|
| Resource |
| Prokaryotes E. coli |
pEZZ18
pEZZ8 |
