期刊
TOXICON
卷 55, 期 8, 页码 1519-1526出版社
PERGAMON-ELSEVIER SCIENCE LTD
DOI: 10.1016/j.toxicon.2010.03.005
关键词
Phage display; Palytoxin; Antibodies; Immunoassay; Shellfish
资金
- Ministry of Education and Science of Spain [AGL2004-08268-C02-01]
- UE [2008-1/003]
- Consolider-Ingenio 2010
Several recombinant antibodies against one of the most potent marine toxins. Palytoxin (PITX, were obtained using two naive human semi-synthetic phage display libraries (Tomlinson I and J) as an effective method for generating specific anti-toxin single-chain variable fragment (scFv) antibodies. After four rounds of panning and selection on free palytoxin adsorbed immunotubes, individual clones were isolated, sequenced and characterized by Enzyme-Linked Immunosorbent Assay (ELISA). Four phage-antibody clones specifically recognized the toxin. A competitive ELISA assay was optimized with one of these phage antibodies giving a very reproducible standard curve with a linear regression (R(2) = 0.9945), showing a working range of 0.0005-500 ng mL(-1). Several spiked shellfish samples were analysed by competitive ELISA to determine the accuracy of the assay, with a mean recovery rate of 90%. This study demonstrates that phage display libraries provide a valuable system for the easy and rapid generation of specific antibody fragments directed against difficult antigenic targets, such as free small molecules. Large-scale, low-cost production of anti-palytoxin scFv antibodies in Escherichia coli (E. coli) is an exciting prospect for the development of rapid and simple detection methods. Our results suggest that anti-palytoxin phage antibodies could be a valuable tool with competitive ELISA to detect palytoxin in natural shellfish samples. (C) 2010 Elsevier Ltd. All rights reserved.
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