Design of Synthetic Peptides for Diagnostics

Meloen, Rob H.; Puijk, Wouter C.; Langeveld, J.P.M.; Langedijk, J.P.M.; Timmerman, Peter

doi:10.2174/1389203033487144

Cited by 31 publications

(30 citation statements)

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“…For research applications, antibodies are by far the most used affinity reagents, including monoclonal antibodies generated by hybridoma technology (30) or monospecific antibodies in which the polyclonal antibody mixture is affinitypurified using synthetic peptides (31) or protein fragments (15). However, the rapid development of new selection methods (29) might make it possible in the future to create renewable affinity reagents in a high throughput manner using in vitro selection principles.…”

Section: Discussionmentioning

confidence: 99%

Antibodypedia, a Portal for Sharing Antibody and Antigen Validation Data

Björling

Uhlén

2008

Molecular & Cellular Proteomics

118

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Antibodies are useful tools to characterize the components of the human proteome and to validate potential protein biomarkers discovered through various clinical proteomics efforts. The lack of validation results across various applications for most antibodies often makes it necessary to perform cumbersome investigations to ensure specificity of a particular antibody in a certain application. A need therefore exists for a standardized system for sharing validation data about publicly available antibodies and to allow antibody providers as well as users to contribute and edit experimental evidence data, including data also on the antigen. Here we describe a new publicly available portal called Antibodypedia, which has been developed to allow sharing of information regarding validation of antibodies in which providers can submit their own validation results and reliability scores. We report standardized validation criteria and submission rules for applications such as Western blots, protein arrays, immunohistochemistry, and immunofluorescence. The contributor is expected to provide experimental evidence and a validation score for each antibody, and the users can subsequently provide feedback and comments on the use of the antibody. The database thus provides a virtual resource of publicly available antibodies toward human proteins with accompanying experimental evidence supporting an individual validation score for each antibody in an application-specific manner.

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Section: Discussionmentioning

confidence: 99%

Antibodypedia, a Portal for Sharing Antibody and Antigen Validation Data

Björling

Uhlén

2008

Molecular & Cellular Proteomics

118

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“…Synthetic peptides usually between 10 and 15 amino acids have also been frequently used to generate affinity reagents (Geysen et al, 1984;Meloen et al, 2003). Synthetic peptides normally consist of a single epitope and therefore subsequent epitope mapping is not needed.…”

Section: Type Of Antigenmentioning

confidence: 99%

“…The problems with cross-reactivity and batch-to-batch variation of polyclonal antibodies have led to the generation of mono-specific antibodies (Olive et al, 2001), in which the polyclonal antibody mixture is affinity purified using the antigen as a ligand. This strategy to generate mono-specific antibodies has been most frequently used for synthetic peptides (Meloen et al, 2003), but recently it has also been extended to antibodies generated by recombinant protein fragments . The name ''mono-specific'' might be somewhat misleading since the affinity-purified antibody mixture often recognizes several epitopes, but the name reflects the fact that only antibodies specific for the antigen, and nothing else, are present in the antibody mixture.…”

Section: Type Of Affinity Reagentmentioning

confidence: 99%

Generation and validation of affinity reagents on a proteome‐wide level

Uhlén

Hober

2008

J of Molecular Recognition

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aThere is a need for protein-specific affinity reagents to explore the gene products encoded by the genome. Recently, systematic efforts to generate validated affinity reagents on a whole human proteome level have been initiated. There are several issues for such efforts, including choice of antigen, type of affinity reagent, and the subsequent validation of the generated protein-specific binders. The advantages and disadvantages with the different approaches are discussed and the problems related to quality assessment of antibodies to be used in multi-platform applications are addressed. This review also describes the efforts to create a virtual resource of validated antibodies using a community-based portal and summarizes the status and visions for the publicly available human protein atlas (www.proteinatlas.org) showing the human protein profiles in a large number of normal and cancer tissues as well as a large set of human cell lines.

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“…These molecules can be constructed from amino-or nucleic acid building blocks. Peptide-like binders with high target binding affinity can be developed by combining library screening, supramolecular chemistry and rational design (Meloen et al, 2003). Their size which is large enough to reproduce a protein binding site, can in some cases adopt a specific three-dimensional structure (Timmerman et al, 2005).…”

Section: Libraries Of Biomoleculesmentioning

confidence: 99%

Fluorescence sensing of intermolecular interactions and development of direct molecular biosensors

2006

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Molecular biosensors are devices of molecular size that are designed for sensing different analytes on the basis of biospecific recognition. They should provide two coupled functions - the recognition (specific binding) of the target and the transduction of information about the recognition event into a measurable signal. The present review highlights the achievements and prospects in design and operation of molecular biosensors for which the transduction mechanism is based on fluorescence. We focus on the general strategy of fluorescent molecular sensing, construction of sensor elements, based on natural and designed biopolymers (proteins and nucleic acids). Particular attention is given to the coupling of sensing elements with fluorescent reporter dyes and to the methods for producing efficient fluorescence responses.

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Design of Synthetic Peptides for Diagnostics

Cited by 31 publications

References 0 publications

Antibodypedia, a Portal for Sharing Antibody and Antigen Validation Data

Antibodypedia, a Portal for Sharing Antibody and Antigen Validation Data

Generation and validation of affinity reagents on a proteome‐wide level

Fluorescence sensing of intermolecular interactions and development of direct molecular biosensors

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