Development and Testing of Recombinant Single Domain Antibodies
Abstract
Conventional antibodies, widely used in detection schemes, such point detectors for biological threats, are large 150 kDa proteins that aggregate irreversibly under harsh conditions such as excessive heating. There is a need for the development of high affinity, highly specific robust recognition elements to overcome the limitations of conventional antibodies. One option is single domain antibodies (sdAb), the molecularly engineered binding elements derived from camelids and shark. SdAb are small 12-16 kDa robust recognition elements able to function after exposure to chemical denaturants and elevated temperatures. These antibody fragments contain only 3 hypervariable loops but can still bind with affinity equivalent to conventional antibody fragments having twice the number of complementarity-determining regions (CDRs) available to interact with the target. We developed and characterized sdAb towards toxins (ricin, abrin and SEB) and viral (filovirus) targets. We employed a combination of strategies, including sandwich immunoassays, surface plasmon resonance, circular dichroism, mutagenesis, and crystallography to help us to understand the mechanism of this bio-molecular recognition and provide critical insights to the basis of the sdAb-target interaction and sdAb properties.
Document Details
- Document Type
- Technical Report
- Publication Date
- Dec 31, 2010
- Accession Number
- ADA545266
Entities
People
- Andrew Hayhurst
- Ellen R Goldman
Organizations
- United States Naval Research Laboratory