Specific Blocking to Improve Biopanning in Biological Samples such as Serum and Hybridoma Supernatants
Screening phage-displayed peptide libraries (biopanning) is an important technique for acquiring peptide ligands and for mapping peptide epitopes recognized by antibodies (Ab). In biological samples, other materials, not only contaminants but also natural constituents, often interfere with biopannin...
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| Format: | Article |
| Language: | English |
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Taylor & Francis Group
2001-04-01
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| Series: | BioTechniques |
| Online Access: | https://www.future-science.com/doi/10.2144/01304st06 |
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| _version_ | 1850153877020606464 |
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| author | Bradley T. Messmer David S. Thaler |
| author_facet | Bradley T. Messmer David S. Thaler |
| author_sort | Bradley T. Messmer |
| collection | DOAJ |
| description | Screening phage-displayed peptide libraries (biopanning) is an important technique for acquiring peptide ligands and for mapping peptide epitopes recognized by antibodies (Ab). In biological samples, other materials, not only contaminants but also natural constituents, often interfere with biopanning. Capture methods use anchoring Abs that reduce the need for purification of the intended panning target from a crude sample. This capture method is analogous to sandwich ELISA. However, when the target molecule concentration is low in the initial mixture, the panning of a captured target often yields epitopes that bind to the capture antibody rather than the target of interest. We have developed a methodology that utilizes specific blocking reagents of the capture Ab to extend the sensitivity and applicability of the capture approach to phage panning. A flowchart is presented to enable the worker to begin panning with the simplest approach and then to employ sandwich capture and specific blocking reagents as necessary. |
| format | Article |
| id | doaj-art-5ac2c1b8dad4423681ffb6331a948d07 |
| institution | OA Journals |
| issn | 0736-6205 1940-9818 |
| language | English |
| publishDate | 2001-04-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | BioTechniques |
| spelling | doaj-art-5ac2c1b8dad4423681ffb6331a948d072025-08-20T02:25:37ZengTaylor & Francis GroupBioTechniques0736-62051940-98182001-04-0130479880210.2144/01304st06Specific Blocking to Improve Biopanning in Biological Samples such as Serum and Hybridoma SupernatantsBradley T. Messmer0David S. Thaler11Rockefeller University, New York, NY, USA1Rockefeller University, New York, NY, USAScreening phage-displayed peptide libraries (biopanning) is an important technique for acquiring peptide ligands and for mapping peptide epitopes recognized by antibodies (Ab). In biological samples, other materials, not only contaminants but also natural constituents, often interfere with biopanning. Capture methods use anchoring Abs that reduce the need for purification of the intended panning target from a crude sample. This capture method is analogous to sandwich ELISA. However, when the target molecule concentration is low in the initial mixture, the panning of a captured target often yields epitopes that bind to the capture antibody rather than the target of interest. We have developed a methodology that utilizes specific blocking reagents of the capture Ab to extend the sensitivity and applicability of the capture approach to phage panning. A flowchart is presented to enable the worker to begin panning with the simplest approach and then to employ sandwich capture and specific blocking reagents as necessary.https://www.future-science.com/doi/10.2144/01304st06 |
| spellingShingle | Bradley T. Messmer David S. Thaler Specific Blocking to Improve Biopanning in Biological Samples such as Serum and Hybridoma Supernatants BioTechniques |
| title | Specific Blocking to Improve Biopanning in Biological Samples such as Serum and Hybridoma Supernatants |
| title_full | Specific Blocking to Improve Biopanning in Biological Samples such as Serum and Hybridoma Supernatants |
| title_fullStr | Specific Blocking to Improve Biopanning in Biological Samples such as Serum and Hybridoma Supernatants |
| title_full_unstemmed | Specific Blocking to Improve Biopanning in Biological Samples such as Serum and Hybridoma Supernatants |
| title_short | Specific Blocking to Improve Biopanning in Biological Samples such as Serum and Hybridoma Supernatants |
| title_sort | specific blocking to improve biopanning in biological samples such as serum and hybridoma supernatants |
| url | https://www.future-science.com/doi/10.2144/01304st06 |
| work_keys_str_mv | AT bradleytmessmer specificblockingtoimprovebiopanninginbiologicalsamplessuchasserumandhybridomasupernatants AT davidsthaler specificblockingtoimprovebiopanninginbiologicalsamplessuchasserumandhybridomasupernatants |