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).