期刊
ELECTROPHORESIS
卷 29, 期 16, 页码 3414-3421出版社
WILEY-V C H VERLAG GMBH
DOI: 10.1002/elps.200800106
关键词
immunocapture; isotachophoresis; magnetic beads; beta-lactoglobulin
资金
- European Community [12793]
- NSFC [NSFC 20375003]
- MOST [MOST NSSP 2006BAF071303]
- EPFL
Protein A-coated magnetic beads (0.3 mu m) have been trapped in a small portion of a neutrally coated capillary (50 mu m id). Anti-beta-lactoglobulin (beta-LG) antibodies have then been immobilized on the beads through strong affinity with protein A to subsequently capture beta-LG from model or real samples. Once the immunocomplexes formed at physiological pH, a discontinuous buffer system has been used to release the partners and preconcentrate them by transient ITP. The antigens and antibodies have finally been separated by CZE and detected by UV absorbance. An LOQ of 55 nM has been achieved. This methodology has been applied to quantify native beta-LG in pasteurized and ultra-high-ternperature-treated bovine milk. All the described procedures, including inimunosorbent preparation, sample extraction, cleanup, preconcentration, and separation are completely automated on a commercial CF instrument. As this CE immunoassay method is simple, rapid, selective, and sensitive, it should be a practical and attractive technology for the analysis of complicated biological samples.
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