Detection and quantification of microcystins (cyanobacterial hepatotoxins) with recombinant antibody fragments isolated from a naïve human phage display library.
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 IC(50) value of 4 microM. 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.
McElhiney
J
J
Department of Molecular and Cell Biology, Institute of Medical Sciences, University of Aberdeen, Foresterhill, UK.
Lawton
L A
LA
Porter
A J
AJ
D000918:Antibody Specificity; D002851:Chromatography, High Pressure Liquid; D003429:Cross Reactions; D000458:Cyanobacteria / Q000737:chemistry*; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007128:Immunoglobulin Fragments / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D008387:Marine Toxins; D052998:Microcystins; D019151:Peptide Library; D010456:Peptides, Cyclic / Q000032:analysis* / Q000276:immunology* / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000276:immunology / Q000302:isolation & purification
D007128:Immunoglobulin Fragments; D008387:Marine Toxins; D052998:Microcystins; D019151:Peptide Library; D010456:Peptides, Cyclic; D011994:Recombinant Proteins; C057862:cyanoginosin LR