Document Detail


Cloning and biological characterization of human single-chain Fv fragments that mediate neutralization of HIV-1.
MedLine Citation:
PMID:  7848589     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
OBJECTIVE: To develop recombinant single-chain Fv fragments against HIV-1 gp120. METHODS: A panel of human monoclonal antibody Fv fragments were generated against the HIV-1 gp120 by affinity selection from an antibody library expressed on the surface of filamentous phage. The library was prepared from peripheral blood lymphocytes of an asymptomatic HIV-1-infected mother with a high neutralization titer. This mother did not transmit HIV-1 to her offspring (non-transmitter). Heavy and light chains were initially amplified separately and combined by splicing by overlap extension to generate Fv fragments. RESULTS: Several clones expressing single-chain Fv fragments bind strongly to HIV-1 gp120 and several were found to neutralize cell-free HIV-1IIIB. Gross epitope mapping suggest that different clones bound to different functional regions on the envelope. The clones also exhibited sequence diversity. CONCLUSIONS: This strategy of cloning resulted in the development of functional human-derived antibody reagents with different anti-HIV-1 biological properties in vitro. These recombinant Fv fragments have potential utility as immune reagents, as well as in the design of potential immunotherapeutics. In addition, these antibody reagents may provide information on the relationship between humoral immunity and maternal-fetal (vertical) HIV-1 transmission.
Authors:
V Srikantan; B Wang; M A Satre; K E Ugen; K Dang; F Scales; A P Godillot; W V Williams; D B Weiner
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  AIDS (London, England)     Volume:  8     ISSN:  0269-9370     ISO Abbreviation:  AIDS     Publication Date:  1994 Nov 
Date Detail:
Created Date:  1995-03-14     Completed Date:  1995-03-14     Revised Date:  2008-11-21    
Medline Journal Info:
Nlm Unique ID:  8710219     Medline TA:  AIDS     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  1525-32     Citation Subset:  IM; X    
Affiliation:
Department of Pathology and Laboratory Medicine, University of Pennsylvania School of Medicine, Philadelphia 19104.
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MeSH Terms
Descriptor/Qualifier:
Acquired Immunodeficiency Syndrome / prevention & control,  transmission*
Amino Acid Sequence
Base Sequence
Cloning, Molecular
DNA Primers
Enzyme-Linked Immunosorbent Assay
Female
HIV Envelope Protein gp120 / immunology*
HIV-1*
Humans
Immunoglobulin Fragments / biosynthesis*,  therapeutic use
Immunoglobulin Heavy Chains / biosynthesis
Immunoglobulin Variable Region / biosynthesis,  therapeutic use
Immunoglobulin kappa-Chains / biosynthesis
Infant, Newborn
Infectious Disease Transmission, Vertical / prevention & control*
Lymphocytes / immunology
Molecular Sequence Data
Neutralization Tests
Polymerase Chain Reaction
Pregnancy
Pregnancy Complications, Infectious / immunology*,  virology
RNA Splicing
Recombinant Proteins / biosynthesis*,  therapeutic use
Chemical
Reg. No./Substance:
0/DNA Primers; 0/HIV Envelope Protein gp120; 0/Immunoglobulin Fragments; 0/Immunoglobulin Heavy Chains; 0/Immunoglobulin Variable Region; 0/Immunoglobulin kappa-Chains; 0/Recombinant Proteins; 0/immunoglobulin Fv

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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