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In situ Protein Detection for Companion Diagnostics.

Gremel G, Grannas K, Sutton LA, Pontén F, Zieba A - Front Oncol (2013)

Bottom Line: The emergence of targeted therapies for cancer has created a need for the development of companion diagnostic tests.We also explore the possibility of using IHC to detect specific protein mutations in addition to DNA-based tests.Finally, we review alternative protein binders and proximity ligation assays and discuss their potential to facilitate the development of novel, targeted therapies against cancer.

View Article: PubMed Central - PubMed

Affiliation: Science for Life Laboratory, Department of Immunology, Genetics and Pathology, Uppsala University , Uppsala , Sweden.

ABSTRACT
The emergence of targeted therapies for cancer has created a need for the development of companion diagnostic tests. Assays developed in recent years are aimed at determining both the effectiveness and safety of specific drugs for a defined group of patients, thus, enabling the more efficient design of clinical trials and also supporting physicians when making treatment-related decisions. Immunohistochemistry (IHC) is a widely accepted method for protein expression analyses in human tissues. Immunohistochemical assays, used to localize and quantitate relative protein expression levels within a morphological context, are frequently used as companion diagnostics during clinical trials and also following drug approval. Herein, we describe established immunochemistry-based methods and their application in routine diagnostics. We also explore the possibility of using IHC to detect specific protein mutations in addition to DNA-based tests. Finally, we review alternative protein binders and proximity ligation assays and discuss their potential to facilitate the development of novel, targeted therapies against cancer.

No MeSH data available.


Related in: MedlinePlus

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Figure 2: Schematic representation of binder formats.

Mentions: In situ affinity-based detection of proteins remains one of the best sources of information about either the healthy status of an individual tissue or potential pathological changes, and is thus applicable within several medical settings. Molecular imaging allows for the early detection and classification of many human diseases and, when specific, permits improved, target-directed therapies. Molecules generated through immunization such as polyclonal, monospecific polyclonal, and monoclonal antibodies continue to be the best established and most widely used binders in diagnostics (90, 91). Methods for the detection of proteins based on antibody recognition often encounter problems due to poor selectivity and/or sensitivity (92). Poorly characterized antibodies and/or insufficient quality control often render them as unsuitable for demanding applications such as companion diagnostics (93). Commercially available antibodies frequently perform very differently within various laboratories and often do not perform as advertised, thus raising doubts regarding their reliability when incorporated into assays requiring high specificity (94). Antibodies can be biochemically and physiologically modified and use of their derivatives, such as single chain variable fragments (scFv) or Fab fragments, may result in the improved detection of a wide range of target molecules (Figure 2) (95). Currently only a few antibodies and recombinant proteins are used within clinical settings, largely due to the reasons outlined above. Recombinant binders that are generated in immune-free, in vitro-based approaches, hold the potential of taking priority over conventional antibodies (96). Stability, specificity, ease of manipulation, low cost, high throughput, and reproducibility of production are some of the advantages that make novel scaffold molecules highly desirable (97). Alternative binders are promising molecules for novel approaches in individualized medicine (Figure 2). They can serve as personalized molecular imaging tools for in vivo, live diagnostics of the changes occurring in the expression of markers following treatment (95).


In situ Protein Detection for Companion Diagnostics.

Gremel G, Grannas K, Sutton LA, Pontén F, Zieba A - Front Oncol (2013)

Schematic representation of binder formats.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3814083&req=5

Figure 2: Schematic representation of binder formats.
Mentions: In situ affinity-based detection of proteins remains one of the best sources of information about either the healthy status of an individual tissue or potential pathological changes, and is thus applicable within several medical settings. Molecular imaging allows for the early detection and classification of many human diseases and, when specific, permits improved, target-directed therapies. Molecules generated through immunization such as polyclonal, monospecific polyclonal, and monoclonal antibodies continue to be the best established and most widely used binders in diagnostics (90, 91). Methods for the detection of proteins based on antibody recognition often encounter problems due to poor selectivity and/or sensitivity (92). Poorly characterized antibodies and/or insufficient quality control often render them as unsuitable for demanding applications such as companion diagnostics (93). Commercially available antibodies frequently perform very differently within various laboratories and often do not perform as advertised, thus raising doubts regarding their reliability when incorporated into assays requiring high specificity (94). Antibodies can be biochemically and physiologically modified and use of their derivatives, such as single chain variable fragments (scFv) or Fab fragments, may result in the improved detection of a wide range of target molecules (Figure 2) (95). Currently only a few antibodies and recombinant proteins are used within clinical settings, largely due to the reasons outlined above. Recombinant binders that are generated in immune-free, in vitro-based approaches, hold the potential of taking priority over conventional antibodies (96). Stability, specificity, ease of manipulation, low cost, high throughput, and reproducibility of production are some of the advantages that make novel scaffold molecules highly desirable (97). Alternative binders are promising molecules for novel approaches in individualized medicine (Figure 2). They can serve as personalized molecular imaging tools for in vivo, live diagnostics of the changes occurring in the expression of markers following treatment (95).

Bottom Line: The emergence of targeted therapies for cancer has created a need for the development of companion diagnostic tests.We also explore the possibility of using IHC to detect specific protein mutations in addition to DNA-based tests.Finally, we review alternative protein binders and proximity ligation assays and discuss their potential to facilitate the development of novel, targeted therapies against cancer.

View Article: PubMed Central - PubMed

Affiliation: Science for Life Laboratory, Department of Immunology, Genetics and Pathology, Uppsala University , Uppsala , Sweden.

ABSTRACT
The emergence of targeted therapies for cancer has created a need for the development of companion diagnostic tests. Assays developed in recent years are aimed at determining both the effectiveness and safety of specific drugs for a defined group of patients, thus, enabling the more efficient design of clinical trials and also supporting physicians when making treatment-related decisions. Immunohistochemistry (IHC) is a widely accepted method for protein expression analyses in human tissues. Immunohistochemical assays, used to localize and quantitate relative protein expression levels within a morphological context, are frequently used as companion diagnostics during clinical trials and also following drug approval. Herein, we describe established immunochemistry-based methods and their application in routine diagnostics. We also explore the possibility of using IHC to detect specific protein mutations in addition to DNA-based tests. Finally, we review alternative protein binders and proximity ligation assays and discuss their potential to facilitate the development of novel, targeted therapies against cancer.

No MeSH data available.


Related in: MedlinePlus