The isolation of novel phage display-derived human recombinant antibodies against CCR5, the major co-receptor of HIV

Moria Shimoni, Alon Herschhorn, Yelena Britan-Rosich, Moshe Kotler, Itai Benhar, Amnon Hizi

Research output: Contribution to journalArticlepeer-review

2 Scopus citations

Abstract

Selecting for antibodies against specific cell-surface proteins is a difficult task due to many unrelated proteins that are expressed on the cell surface. Here, we describe a method to screen antibody-presenting phage libraries against native cell-surface proteins. We applied this method to isolate antibodies that selectively recognize CCR5, which is the major co-receptor for HIV entry (consequently, playing a pivotal role in HIV transmission and pathogenesis). We employed a phage screening strategy by using cells that co-express GFP and CCR5, along with an excess of control cells that do not express these proteins (and are otherwise identical to the CCR5-expressing cells). These control cells are intended to remove most of the phages that bind the cells nonspecifically; thus leading to an enrichment of the phages presenting anti-CCR5-specific antibodies. Subsequently, the CCR5-presenting cells were quantitatively sorted by flow cytometry, and the bound phages were eluted, amplified, and used for further successive selection rounds. Several different clones of human single-chain Fv antibodies that interact with CCR5-expressing cells were identified. The most specific monoclonal antibody was converted to a full-length IgG and bound the second extracellular loop of CCR5. The experimental approach presented herein for screening for CCR5-specific antibodies can be applicable to screen antibody-presenting phage libraries against any cell-surface expressed protein of interest.

Original languageEnglish (US)
Pages (from-to)277-290
Number of pages14
JournalViral Immunology
Volume26
Issue number4
DOIs
StatePublished - Aug 1 2013

Bibliographical note

Copyright:
Copyright 2014 Elsevier B.V., All rights reserved.

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