Development and Testing of Recombinant Single Domain Antibodies

reportActive / Technical Report | Accession Number: ADA545266 | Open PDF

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.

Author(s):
Goldman, Ellen ; Hayhurst, Andrew
Author Organization(s):
NAVAL RESEARCH LAB WASHINGTON DC
Descriptive Note:
Final rept. 1 Apr 2006-30 Jun 2010
Supplementary Note:
The original document contains color images. DOI: 10.21236/ADA545266
Pagination:
0016

Security Markings

DOCUMENT & CONTEXTUAL SUMMARY

Distribution:
Approved For Public Release
Distribution Statement:
Approved For Public Release; Distribution Is Unlimited.

RECORD

Collection: TR
Identifying Numbers
Report Number(s):
ARO-50904-CH.1
Monitor Series:
50904-CH.1, ARO
 Subject Terms
Joint Capability Areas:
JCA_5_Command and Control; JCA_5.3_Planning; JCA_5.3.4_Develop Courses of Action; JCA_5.3.5_Analyze Courses of Action; JCA_5.3.3_Develop Strategy; JCA_1.2.3_Educating; JCA_4.6_Engineering
Communities of Interest:
No COI(s) Identified
Descriptor(s):
*BIOLOGICAL DETECTION, *ANTIBODIES, BIOLOGICAL AGENT DETECTORS, IMMUNOASSAY, RICIN, RECOGNITION
Field(s)/Group(s):
Biochemistry, Chemical, Biological and Radiological Warfare, Miscellaneous Detection and Detectors
Keyword(s):
FILOVIRUS, SINGLE DOMAIN ANTIBODIES, PE106013
Report Date:
2010 Dec 31
Creation Date:
2011 Aug 01