Using Phage and Yeast Display to Select Hundreds of Monoclonal Antibodies: Application to Antigen 85, a Tuberculosis Biomarker

Ferrara, Fortunato; Naranjo, Leslie A.; Kumar, Sandeep; Gaiotto, Tiziano; Mukundan, Harshini; Swanson, Basil; Bradbury, Andrew R.  M.

doi:10.1371/journal.pone.0049535

Using Phage and Yeast Display to Select Hundreds of Monoclonal Antibodies: Application to Antigen 85, a Tuberculosis Biomarker

Journal Article · Wed Nov 14 00:00:00 EST 2012 · PLoS ONE

DOI:https://doi.org/10.1371/journal.pone.0049535· OSTI ID:1627561

Ferrara, Fortunato ^[1]; Naranjo, Leslie A. ^[2]; Kumar, Sandeep ^[2]; Gaiotto, Tiziano ^[2]; Mukundan, Harshini ^[2]; Swanson, Basil ^[2]; Bradbury, Andrew R. M. ^[2]

Los Alamos National Lab. (LANL), Los Alamos, NM (United States); DOE/OSTI
Los Alamos National Lab. (LANL), Los Alamos, NM (United States)

Background: Current diagnostic methods for tuberculosis (TB), a major global health challenge that kills nearly two million people annually, are time-consuming and inadequate. During infection a number of bacterial molecules that play a role in the infective process are released and have been proposed as biomarkers for early TB diagnosis. Antigen 85 (Ag85) is the most abundant secreted TB protein, and a potential target for this diagnostic approach. One of the bottlenecks in the direct detection of such bacterial targets is the availability of robust, sensitive, specific antibodies. Methods: Using Ag85 as a model, we describe a method to select antibodies against any potential target using a novel combination of phage and yeast display that exploits the advantage of each approach. Results: The efficiency of this approach was attested to by the 111 specific antibodies identified in initial screens. These were assessed for binding to the different Ag85 subunits, affinity, and activity in sandwich assays. Conclusions: The novelty of this approach lies in the possibility of screening the entire output of a phage antibody selection in a single experiment by yeast display. This can be considered analogous to carrying out a million ELISAs. The monoclonal antibodies (mAbs) identified in this way show high binding affinity and selectivity for the antigens and offer an advantage over traditional mAbs produced by relatively expensive and time consuming techniques. This approach has wide applicability, and the affinity of selected antibodies can be significantly improved, if required.

View Accepted Manuscript (DOE)

Research Organization:: Los Alamos National Laboratory (LANL), Los Alamos, NM (United States)

Sponsoring Organization:: USDOE Laboratory Directed Research and Development (LDRD) Program; USDOE Office of Science (SC), Biological and Environmental Research (BER). Biological Systems Science Division

OSTI ID:: 1627561

Journal Information:: PLoS ONE, Journal Name: PLoS ONE Journal Issue: 11 Vol. 7; ISSN 1932-6203

Publisher:: Public Library of ScienceCopyright Statement

Country of Publication:: United States

Language:: English

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Using Phage and Yeast Display to Select Hundreds of Monoclonal Antibodies: Application to Antigen 85, a Tuberculosis Biomarker

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