Method for quantitating trace amounts of an analyte in a sample by affinity capillary electrophoresis
US5348633A · kind A · utility
Assignee
Inventors
Key dates
| Filing date | Jan 22, 1993 |
| Grant date | Sep 20, 1994 |
| Priority date | — |
| Expiry date | Jan 22, 2013 |
Classification
- Technology area (CPC G)Physics
- CPC primaryG01N33/6857
- WIPO fieldMeasurement
- WIPO sectorInstruments
Abstract
A method for quantitative detection of trace amounts of an analyte in a sample is disclosed. The method in the preferred embodiment includes providing an Fab' fragment of an immunoglobulin labelled at a reactive sulfhydryl group with a fluorescent dye; combining the labelled Fab' fragment with a sample that may contain the analyte; concentrating the elements of the resulting mixture in an electric field; separating the labelled analyte/agent complex formed from any unreacted labelled agent using capillary electrophoretic methods; and detecting the fluorescent signal of the labelled analyte/agent complex. The invention also is directed to a method of producing a labelled Fab' fragment that includes cleaving an immunoglobulin molecule to obtain one F(ab').sub.2 fragment; reducing the disulfide-bonds of the F(ab').sub.2 fragment to obtain two Fab' fragments each having at least one free, reactive sulfhydryl group; and mixing an Fab' fragment having at least one free sulfhydryl group with a fluorescent dye reactive with the free sulfhydryl to form a labelled Fab' fragment. Preferably, prior to the final mixing step, intrastrand disulfide bonds are formed by oxidation within each Fab' f…
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.