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Titolo:
LIPID-TAGGED ANTIBODIES - BACTERIAL EXPRESSION AND CHARACTERIZATION OF A LIPOPROTEIN SINGLE-CHAIN ANTIBODY FUSION PROTEIN
Autore:
LAUKKANEN ML; TEERI TT; KEINANEN K;
Indirizzi:
VTT BIOTECH LAB,POB 202 SF-02151 ESPOO FINLAND VTT BIOTECH LAB,POB 202 SF-02151 ESPOO FINLAND
Titolo Testata:
Protein engineering
fascicolo: 4, volume: 6, anno: 1993,
pagine: 449 - 454
SICI:
0269-2139(1993)6:4<449:LA-BEA>2.0.ZU;2-L
Fonte:
ISI
Lingua:
ENG
Soggetto:
ESCHERICHIA-COLI; OUTER-MEMBRANE; LIPOSOMES; PHAGE; LOCALIZATION; LIBRARIES; FRAGMENT; CELLS; MOUSE; DNA;
Keywords:
ANTIBODY ENGINEERING; EXPRESSION IN ESCHERICHIA-COLI; FUSION PROTEIN; LIPOPROTEIN; TRITON X-114;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Citazioni:
30
Recensione:
Indirizzi per estratti:
Citazione:
M.L. Laukkanen et al., "LIPID-TAGGED ANTIBODIES - BACTERIAL EXPRESSION AND CHARACTERIZATION OF A LIPOPROTEIN SINGLE-CHAIN ANTIBODY FUSION PROTEIN", Protein engineering, 6(4), 1993, pp. 449-454

Abstract

In order to achieve a stable and functional immobilization of antibodies, we investigated the possibility of adding hydrophobic membrane anchors to antibody fragments expressed in Escherichia coli. The DNA sequence encoding the signal peptide and the nine N-terminal amino acid residues of the major lipoprotein of E.coli was fused to the sequence of an anti-2-phenyloxazolone single-chain Fv antibody fragment [Takkinen et al. (1991) Protein Engng, 4, 837-841]. The expression of the fusion construct in E.coli resulted in specific accumulation of an immunoreactive 28 kDa polypeptide. Unlike the unmodified single-chain Fv fragment, the fusion protein was cell-associated, labelled by [H-3]palmitate which is indicative of the presence of N-terminal lipid modification, partitioned into the detergent phase upon Triton X-114 phase separation and was localized predominantly in the bacterial outer membrane. The fusion antibody displayed specific 2-phenyloxazolone-binding activity in the membrane-bound form and after solubilization with non-ionicdetergents. Furthermore, upon removal of detergent the fusion antibody was incorporated into proteoliposomes which displayed specific hapten-binding activity. Our results show that antibodies can be converted to membrane-bound proteins with retention of antigen-binding properties by introduction of lipid anchors during biosynthesis. This approach may prove useful in the design of immunoliposomes and immunosensors.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 31/03/20 alle ore 05:15:16