Use of intramolecularly, covalently cross-linked proteins as binding partners in immunoassays
AssigneeRoche Diagnostic Operations, Inc.
InventorsBarbara Upmeier, Dittmar Schlieper, Frederic Donie
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Abstract
The invention concerns the use of intramolecularly, covalently cross-linked proteins and covalently cross-linked reverse transcriptase from HIV as immunological binding partners in immunoassays. It also concerns immunological test procedures for detecting an analyte in a sample in which intramolecularly, covalently cross-linked proteins are used as binding partners, and it further concerns intramolecularly, covalently cross-linked reverse transcriptase from HIV and a method for producing this reverse transcriptase.
Claims12
A method for detecting an analyte in a sample comprising the steps of:
- a. combining the sample with an intramolecularly, covalently cross-linked HIV reverse transcriptase in the absence of detectable intermolecular multimers of the HIV reverse transcriptase, wherein the HIV reverse transcriptase specifically binds with the analyte or with a substance bound to the analyte to form a complex,
- b. adding to the combination formed in step (a) a binding partner provided with a label that combines with the complex formed in step (a) to produce a detectable signal, and
- c. determining the signal produced in step (b) as a measure of the analyte in the sample.
A method for detecting an analyte in a sample comprising the steps of:
- a. contacting the sample with a composition comprising an intramolecularly, covalently cross-linked HIV reverse transcriptase, wherein said composition comprises less than about 5% of total intermolecular multimers of HIV reverse transcriptase relative to the eluted peak intramolecularly, covalently cross-linked HIV reverse transcriptase as determined in a gel permeation chromatograph, wherein the intramolecularly, covalently cross-linked HIV reverse transcriptase specifically binds with the analyte or with a substance bound to the analyte to form a complex,
- b. adding to the combination formed in step (a) a binding partner provided with a label that combines with the complex formed in step (a) to produce a detectable signal, and
- c. determining the signal produced in step (b) as a measure of the analyte in the sample.
Dependencies are inferred from the claim text.