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1994-07-02
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Document 0095
DOCN M9470095
TI Recombinant single-chain antibody peptide conjugates expressed in
Escherichia coli for the rapid diagnosis of HIV.
DT 9409
AU Lilley GG; Dolezal O; Hillyard CJ; Bernard C; Hudson PJ; CSIRO Division
of Biomolecular Engineering, Victoria, Australia.
SO J Immunol Methods. 1994 May 16;171(2):211-26. Unique Identifier :
AIDSLINE MED/94253566
AB Recombinant single chain Fv (scFv) antibody fragments can form the basis
of a rapid, whole-blood diagnostic assay. The scFv described in this
study is derived from a monoclonal antibody which has a high affinity
for glycophorin A, an abundant glycoprotein on the human red blood cell
membrane surface. The prototype reagent built around the scFv was
designed to detect, in whole blood samples, the presence of antibodies
that have arisen through infection with a foreign organism such as human
immunodeficiency virus. The scFv was composed of the antibody
heavy-chain variable domain (Vh) joined by a 15 residue linker (GGGGS)3-
to the light-chain variable domain (V1) terminated by either a
C-terminal octapeptide tail (FLAG) or a 35 amino acid segment from the
gp41 surface glycoprotein of HIV-1. Constructs were cloned into a
Escherichia coli expression vector, pHFA, and expressed in a soluble
form into culture supernatant. The product retained anti-glycophorin
activity which could be detected directly in culture supernatants by
ELISA. Furthermore, the scFv-epitope fusion functioned efficiently in
the whole blood agglutination assay and was able to distinguish between
HIV-1 positive and negative sera.
DE Amino Acid Sequence Antibodies, Monoclonal/*BIOSYNTHESIS/DIAGNOSTIC
USE/*GENETICS Antigen-Antibody Complex/*BIOSYNTHESIS/DIAGNOSTIC
USE/GENETICS Antigenic Determinants/GENETICS/ISOLATION & PURIF Base
Sequence Cloning, Molecular Comparative Study Escherichia
coli/*GENETICS/*METABOLISM Human HIV Infections/BLOOD/*DIAGNOSIS
HIV-1 Immunoglobulin Fragments/*BIOSYNTHESIS/DIAGNOSTIC USE/*GENETICS
Molecular Sequence Data Peptides/BIOSYNTHESIS/DIAGNOSTIC USE/GENETICS
Recombinant Fusion Proteins/*BIOSYNTHESIS/DIAGNOSTIC USE/ *GENETICS
Support, Non-U.S. Gov't JOURNAL ARTICLE
SOURCE: National Library of Medicine. NOTICE: This material may be
protected by Copyright Law (Title 17, U.S.Code).