Improving the biophysical properties of anti-ricin single-domain antibodies
Single-domain antibodies (sdAbs) derived from heavy-chain only antibodies produced in camelids are attractive immunoreagents due to their small size, high affinity, and ability to refold and retain binding activity after denaturation. It has been observed that some sdAbs, however, exhibit undesirabl...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2015-06-01
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| Series: | Biotechnology Reports |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2215017X15000041 |
| _version_ | 1818506095183790080 |
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| author | Kendrick B. Turner Jinny L. Liu Dan Zabetakis Audrey Brozozog Lee George P. Anderson Ellen R. Goldman |
| author_facet | Kendrick B. Turner Jinny L. Liu Dan Zabetakis Audrey Brozozog Lee George P. Anderson Ellen R. Goldman |
| author_sort | Kendrick B. Turner |
| collection | DOAJ |
| description | Single-domain antibodies (sdAbs) derived from heavy-chain only antibodies produced in camelids are attractive immunoreagents due to their small size, high affinity, and ability to refold and retain binding activity after denaturation. It has been observed that some sdAbs, however, exhibit undesirable properties including reduced solubility when subjected to heating or upon long-term storage at production-relevant concentrations, which can limit their usefulness. Using a multi-step, rational design approach that included consensus-sequence driven sequence repairs, the alteration of net protein charge, and the introduction of non-native disulfide bonds, augmented solubility and increased melting temperatures were achieved. The improved sdAbs tolerated storage in solution at high concentration (10 mg/mL) and were able to withstand multiple cycles of heating to high temperature (70 °C). This work demonstrates a pathway for improving the biophysical characteristics of sdAbs which is essential for expanding their utility for both diagnostic as well as therapeutic applications. |
| first_indexed | 2024-12-10T21:59:53Z |
| format | Article |
| id | doaj.art-df19dd9271cc41cfad1cda45c997dd8b |
| institution | Directory Open Access Journal |
| issn | 2215-017X |
| language | English |
| last_indexed | 2024-12-10T21:59:53Z |
| publishDate | 2015-06-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Biotechnology Reports |
| spelling | doaj.art-df19dd9271cc41cfad1cda45c997dd8b2022-12-22T01:31:56ZengElsevierBiotechnology Reports2215-017X2015-06-016C273510.1016/j.btre.2015.01.001Improving the biophysical properties of anti-ricin single-domain antibodiesKendrick B. Turner0Jinny L. Liu1Dan Zabetakis2Audrey Brozozog Lee3George P. Anderson4Ellen R. Goldman5American Society for Engineering Education, Postdoctoral Fellow at the Naval Research Laboratory, Washington, DC 20375, USACenter for Biomolecular Science and Engineering, Naval Research Laboratory, Washington, DC 20375, USACenter for Biomolecular Science and Engineering, Naval Research Laboratory, Washington, DC 20375, USANOVA Research Inc., 1900 Elkin St. Suite 230, Alexandria, VA 22308, USACenter for Biomolecular Science and Engineering, Naval Research Laboratory, Washington, DC 20375, USACenter for Biomolecular Science and Engineering, Naval Research Laboratory, Washington, DC 20375, USASingle-domain antibodies (sdAbs) derived from heavy-chain only antibodies produced in camelids are attractive immunoreagents due to their small size, high affinity, and ability to refold and retain binding activity after denaturation. It has been observed that some sdAbs, however, exhibit undesirable properties including reduced solubility when subjected to heating or upon long-term storage at production-relevant concentrations, which can limit their usefulness. Using a multi-step, rational design approach that included consensus-sequence driven sequence repairs, the alteration of net protein charge, and the introduction of non-native disulfide bonds, augmented solubility and increased melting temperatures were achieved. The improved sdAbs tolerated storage in solution at high concentration (10 mg/mL) and were able to withstand multiple cycles of heating to high temperature (70 °C). This work demonstrates a pathway for improving the biophysical characteristics of sdAbs which is essential for expanding their utility for both diagnostic as well as therapeutic applications.http://www.sciencedirect.com/science/article/pii/S2215017X15000041ThermostabilityCircular dichroismCamelid |
| spellingShingle | Kendrick B. Turner Jinny L. Liu Dan Zabetakis Audrey Brozozog Lee George P. Anderson Ellen R. Goldman Improving the biophysical properties of anti-ricin single-domain antibodies Biotechnology Reports Thermostability Circular dichroism Camelid |
| title | Improving the biophysical properties of anti-ricin single-domain antibodies |
| title_full | Improving the biophysical properties of anti-ricin single-domain antibodies |
| title_fullStr | Improving the biophysical properties of anti-ricin single-domain antibodies |
| title_full_unstemmed | Improving the biophysical properties of anti-ricin single-domain antibodies |
| title_short | Improving the biophysical properties of anti-ricin single-domain antibodies |
| title_sort | improving the biophysical properties of anti ricin single domain antibodies |
| topic | Thermostability Circular dichroism Camelid |
| url | http://www.sciencedirect.com/science/article/pii/S2215017X15000041 |
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