Detection and quantification of microcystins (cyanobacterial hepatotoxins) with recombinant antibody fragments isolated from a naive human phage display library

J McElhiney, L A Lawton, A J R Porter

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33 Citations (Scopus)

Abstract

Single-chain antibody fragments against the cyanobacterial hepatotoxin microcystin-LR were isolated from a naive human phage display library and expressed in Escherichia coli. In competition enzyme-linked immunosorbent assay (ELISA), the most sensitive antibody clone selected from the library detected free microcystin-LR with an IC50 value of 4 muM. It was found to cross react with three other microcystin variants - microcystin-RR, microcystin-LW and microcystin-LF - and detected microcystins in extracts of the cyanobacterium Microcystis aeruginosa, found to contain the toxins by high-performance liquid chromatography (HPLC). The quantification of microcystins in these extracts by ELISA and HPLC showed good correlation. Although the antibody isolated in this study was considerably less sensitive than the polyclonal and monoclonal antibodies already available for microcystin detection, phage display technology represents a cheaper, more rapid alternative for the production of anti-microcystin antibodies than the methods currently in use. (C) 2000 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved.

Original languageEnglish
Pages (from-to)83-88
Number of pages6
JournalFEMS Microbiology Letters
Volume193
Publication statusPublished - 2000

Keywords

  • cyanobacteria
  • microcystin
  • antibody fragment
  • immunoassay
  • ESCHERICHIA-COLI
  • PURIFICATION
  • CHROMATOGRAPHY
  • PHOSPHATASE-1
  • SELECTION
  • LR

Cite this

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title = "Detection and quantification of microcystins (cyanobacterial hepatotoxins) with recombinant antibody fragments isolated from a naive human phage display library",
abstract = "Single-chain antibody fragments against the cyanobacterial hepatotoxin microcystin-LR were isolated from a naive human phage display library and expressed in Escherichia coli. In competition enzyme-linked immunosorbent assay (ELISA), the most sensitive antibody clone selected from the library detected free microcystin-LR with an IC50 value of 4 muM. It was found to cross react with three other microcystin variants - microcystin-RR, microcystin-LW and microcystin-LF - and detected microcystins in extracts of the cyanobacterium Microcystis aeruginosa, found to contain the toxins by high-performance liquid chromatography (HPLC). The quantification of microcystins in these extracts by ELISA and HPLC showed good correlation. Although the antibody isolated in this study was considerably less sensitive than the polyclonal and monoclonal antibodies already available for microcystin detection, phage display technology represents a cheaper, more rapid alternative for the production of anti-microcystin antibodies than the methods currently in use. (C) 2000 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved.",
keywords = "cyanobacteria, microcystin, antibody fragment, immunoassay, ESCHERICHIA-COLI, PURIFICATION, CHROMATOGRAPHY, PHOSPHATASE-1, SELECTION, LR",
author = "J McElhiney and Lawton, {L A} and Porter, {A J R}",
year = "2000",
language = "English",
volume = "193",
pages = "83--88",
journal = "FEMS Microbiology Letters",
issn = "0378-1097",
publisher = "Oxford University Press",

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TY - JOUR

T1 - Detection and quantification of microcystins (cyanobacterial hepatotoxins) with recombinant antibody fragments isolated from a naive human phage display library

AU - McElhiney, J

AU - Lawton, L A

AU - Porter, A J R

PY - 2000

Y1 - 2000

N2 - Single-chain antibody fragments against the cyanobacterial hepatotoxin microcystin-LR were isolated from a naive human phage display library and expressed in Escherichia coli. In competition enzyme-linked immunosorbent assay (ELISA), the most sensitive antibody clone selected from the library detected free microcystin-LR with an IC50 value of 4 muM. It was found to cross react with three other microcystin variants - microcystin-RR, microcystin-LW and microcystin-LF - and detected microcystins in extracts of the cyanobacterium Microcystis aeruginosa, found to contain the toxins by high-performance liquid chromatography (HPLC). The quantification of microcystins in these extracts by ELISA and HPLC showed good correlation. Although the antibody isolated in this study was considerably less sensitive than the polyclonal and monoclonal antibodies already available for microcystin detection, phage display technology represents a cheaper, more rapid alternative for the production of anti-microcystin antibodies than the methods currently in use. (C) 2000 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved.

AB - Single-chain antibody fragments against the cyanobacterial hepatotoxin microcystin-LR were isolated from a naive human phage display library and expressed in Escherichia coli. In competition enzyme-linked immunosorbent assay (ELISA), the most sensitive antibody clone selected from the library detected free microcystin-LR with an IC50 value of 4 muM. It was found to cross react with three other microcystin variants - microcystin-RR, microcystin-LW and microcystin-LF - and detected microcystins in extracts of the cyanobacterium Microcystis aeruginosa, found to contain the toxins by high-performance liquid chromatography (HPLC). The quantification of microcystins in these extracts by ELISA and HPLC showed good correlation. Although the antibody isolated in this study was considerably less sensitive than the polyclonal and monoclonal antibodies already available for microcystin detection, phage display technology represents a cheaper, more rapid alternative for the production of anti-microcystin antibodies than the methods currently in use. (C) 2000 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved.

KW - cyanobacteria

KW - microcystin

KW - antibody fragment

KW - immunoassay

KW - ESCHERICHIA-COLI

KW - PURIFICATION

KW - CHROMATOGRAPHY

KW - PHOSPHATASE-1

KW - SELECTION

KW - LR

M3 - Article

VL - 193

SP - 83

EP - 88

JO - FEMS Microbiology Letters

JF - FEMS Microbiology Letters

SN - 0378-1097

ER -