US 2022/0011301 A1 · STREPTAVIDIN-COATED SOLID PHASES WITH A MEMBER OF A BINDING PAIR

Abstract

The present disclosure relates to a solid phase coated with (strept)avidin and having attached thereto, by way of biotin:(strept)avidin interaction, a biotinylated first member of a binding pair, wherein the attached first member is capable of binding to a second member of the binding pair, but is not capable of binding to biotin or to (strept)avidin, and wherein no member of the binding pair is capable of hybridizing with a naturally-occurring single-stranded nucleic acid. The solid phase is particularly useful in immunoassays with samples having high content in biotin or (strept)avidin-binding derivatives thereof. The present disclosure further provides uses, kits and methods, particularly for determination of an analyte in a sample.

Claims25

Dependent claims

A solid phase coated with (strept)avidin and having attached thereto, by way of <biotin:(strept)avidin> interaction, a biotinylated first member of a binding pair, wherein the attached first member is capable of binding to a second member of the binding pair, but is not capable of binding to biotin or to (strept)avidin, wherein the second member is capable of becoming bound by the first member when the second member is part of a conjugate comprising any of an analyte, an analyte analogon, and an analyte-specific capturing agent, and wherein no member of the binding pair is capable of hybridizing with a naturally-occurring single-stranded nucleic acid.

A method of preparing a solid phase having attached thereto a member of a binding pair, the method comprising the steps of

(a) providing a solid phase coated with (strept)avidin;
(b) selecting a binding pair with a first member and a second member;
(c) providing the first member of the binding pair selected in step (b);
(d) biotinylating the first member of step (c);
(e) attaching the biotinylated first member obtained from step (d) to the solid phase of step (a) by contacting the biotinylated first member with the coated solid phase and incubating, thereby attaching the biotinylated first member to the solid phase by way of biotin-(strept)avidin interaction;
wherein in step (b) the pair is selected such that
without biotinylation the first and the second member of the binding pair are not capable of binding to streptavidin,
in biotinylated form and non-covalently attached to the coated solid phase by way of a biotin:(strept)avidin bond the first member of the binding pair is capable of binding to the second member,
in conjugated form and covalently attached to an analyte-specific capturing agent the second member of the binding pair is capable of binding to the biotinylated first member attached to the solid phase, and
no member of the binding pair is capable of hybridizing with a naturally-occurring single-stranded nucleic acid;
thereby obtaining the solid phase having attached thereto the member of the binding pair.

8 more elements

Dependencies are inferred from the claim text.

Streptavidin-coated solid phases with a member of a binding pair

Life of this patent

Streptavidin-coated solid phases with a member of a binding pair

Life of this patent

Abstract

Claims25