Use of a gene-targeted phage display random epitope library to map an antigenic determinant on the bluetongue virus outer capsid protein VP5

Wang, Lin‐Fa; Plessis, D.H. Du; White, John R.; Hyatt, Alex D.; Eaton, Bryan T.

doi:10.1016/0022-1759(94)00235-o

Cited by 78 publications

(39 citation statements)

References 39 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…The selected phage clones were amplified to a high titer and purified twice by precipitation with 20% (wt/vol) polyethylene glycol 8000 (PEG 8000)-2.5 M NaCl according to the method described by Wang et al (23). The phage titration method was adapted from Sambrook et al (17).…”

Section: Bacterial Strains and Reagents Escherichia Colimentioning

confidence: 99%

Mimotopes of the Vi Antigen ofSalmonella entericaSerovar Typhi Identified from Phage Display Peptide Library

Tang

Tan

Devi

et al. 2003

Clin Vaccine Immunol

Self Cite

View full text Add to dashboard Cite

The capsular polysaccharide Vi antigen (ViCPS) is an essential virulence factor and also a protective antigen of Salmonella enterica serovar Typhi. A random 12-mer phage-displayed peptide library was used to identify mimotopes (epitope analogues) of this antigen by panning against a ViCPS-specific monoclonal antibody (MAb) ATVi. Approximately 75% of the phage clones selected in the fourth round carried the peptide sequence TSHHDSHGLHRV, and the rest of the clones harbored ENHSPVNIAHKL and other related sequences. These two sequences were also obtained in a similar panning process by using pooled sera from patients with a confirmed diagnosis of typhoid fever, suggesting they mimic immunodominant epitopes of ViCPS antigens. Binding of MAb ATVi to the mimotopes was specifically blocked by ViCPS, indicating that they interact with the same binding site (paratope) of the MAb. Data and reagents generated in this study have important implications for the development of peptide-base diagnostic tests and peptide vaccines and may also provide a better understanding of the pathogenesis of typhoid fever.

show abstract

Section: Bacterial Strains and Reagents Escherichia Colimentioning

confidence: 99%

Mimotopes of the Vi Antigen ofSalmonella entericaSerovar Typhi Identified from Phage Display Peptide Library

Tang

Tan

Devi

et al. 2003

Clin Vaccine Immunol

Self Cite

View full text Add to dashboard Cite

show abstract

“…The BOT1-selected S25 and 3D12 clones are larger (500 to 600 bp) than those in the library (150 to 300 bp). In contrast, other gene fragment selections (50 to 400 bp) from multivalent rather than monovalent display libraries yield small epitopes (i.e., 50 to 200 bp) (3,4,8,9,11,20,27) that are perhaps related to multivalent, smaller fragments with higher functional affinity.…”

mentioning

confidence: 93%

Epitope Mapping of Neutralizing Botulinum Neurotoxin A Antibodies by Phage Display

2001

View full text Add to dashboard Cite

show abstract

“…Although traditional phage display has been successfully applied to gene rich bacterial genomes (Jacobsson and Frykberg 1995Jacobsson et al 1997) and individual genes (Parmley and Smith 1989;Du Plessis et al 1995;Petersen et al 1995;Wang et al 1995;Bluthner et al 1996Bluthner et al , 1999, to identify antibody epitopes or binding partners, it suffers from the problem that only one clone in 18, if starting with DNA encoding an ORF, will be correctly in frame (one clone in three will start correctly, one clone in three will end correctly, and one clone in two will have the correct orientation), although experiments with synthetic peptide libraries have indicated that stop codons do not necessarily prevent display (Carcamo et al 1998). Although this high rate of nonfunctional inserts may be tolerable when starting with DNA from a single gene or even a small gene rich genome, in which complete functional representation can be obtained with relatively small libraries, it may become impractical if using more complex DNA sources.…”

mentioning

confidence: 99%

Selecting Open Reading Frames From DNA

Zacchi

Sblattero²,

Florian³

et al. 2003

Genome Res.

View full text Add to dashboard Cite

We describe a method to select DNA encoding functional open reading frames (ORFs) from noncoding DNA within the context of a specific vector. Phage display has been used as an example, but any system requiring DNA encoding protein fragments, for example, the yeast two-hybrid system, could be used. By cloning DNA fragments upstream of a fusion gene, consisting of the β-lactamase gene flanked by lox recombination sites, which is, in turn, upstream of gene 3 from fd phage, only those clones containing DNA fragments encoding ORFs confer ampicillin resistance and survive. After selection, the β-lactamase gene can be removed by Cre recombinase, leaving a standard phage display vector with ORFs fused to gene 3. This vector has been tested on a plasmid containing tissue transglutaminase. All surviving clones analyzed by sequencing were found to contain ORFs, of which 83% were localized to known genes, and at least 80% produced immunologically detectable polypeptides. Use of a specific anti-tTG monoclonal antibody allowed the identification of clones containing the correct epitope. This approach could be applicable to the efficient selection of random ORFs representing the coding potential of whole organisms, and their subsequent downstream use in a number of different systems

show abstract

Use of a gene-targeted phage display random epitope library to map an antigenic determinant on the bluetongue virus outer capsid protein VP5

Cited by 78 publications

References 39 publications

Mimotopes of the Vi Antigen ofSalmonella entericaSerovar Typhi Identified from Phage Display Peptide Library

Mimotopes of the Vi Antigen ofSalmonella entericaSerovar Typhi Identified from Phage Display Peptide Library

Epitope Mapping of Neutralizing Botulinum Neurotoxin A Antibodies by Phage Display

Selecting Open Reading Frames From DNA

Contact Info

Product

Resources

About