Abstract：
BACKGROUND:Shark heavy chain antibody, also called new antigen receptor (NAR), consists of one single Variable domain (VH), containing only two complementarity-determining regions (CDRs). The antigen binding affinity and specificity are mainly determined by these two CDRs. The good solubility, excellent thermal stability and complex sequence variation of small single domain antibodies (sdAbs) make them attractive alternatives to conventional antibodies. In this report, we construct and characterize a diversity enhanced semi-synthetic NAR V display library based on naturally occurring NAR V sequences. RESULTS:A semi-synthetic shark sdAb display library with a complexity close to 1e9 was constructed. This was achieved by introducing size and sequence variations in CDR3 using randomized CDR3 primers of three different lengths. Binders against three toxins, staphylococcal enterotoxin B (SEB), ricin, and botulinum toxin A (BoNT/A) complex toxoid, were isolated from panning the display library. Soluble sdAbs from selected binders were purified and evaluated using direct binding and thermal stability assays on the Luminex 100. In addition, sandwich assays using sdAb as the reporter element were developed to demonstrate their utility for future sensor applications. CONCLUSION:We demonstrated the utility of a newly created hyper diversified shark NAR displayed library to serve as a source of thermal stable sdAbs against a variety of toxins.
journal_name
BMC Biotechnoljournal_title
BMC biotechnologyauthors
Liu JL,Anderson GP,Goldman ERdoi
10.1186/1472-6750-7-78subject
Has Abstractpub_date
2007-11-19 00:00:00pages
78issn
1472-6750pii
1472-6750-7-78journal_volume
7pub_type
杂志文章abstract：BACKGROUND:Analysis of protein-protein interactions (PPIs) is a valuable approach for the characterization of huge networks of protein complexes or proteins of unknown function. Co-immunoprecipitation (coIP) using affinity resins coupled to protein A/G is the most widely used method for PPI detection. However, this tra...
journal_title：BMC biotechnology
pub_type： 杂志文章
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更新日期：2008-03-05 00:00:00
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pub_type： 杂志文章,收录出版
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pub_type： 杂志文章
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