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Bis-biotinylation tags

A dual biotin, biotinylation technology, applied in biological testing, chemical instruments and methods, peptides, etc., can solve cumbersome and other problems

Active Publication Date: 2016-02-03
PACIFIC BIOSCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, Howarth's method involves mixing 2 recombinant streptavidins and is tedious

Method used

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Embodiment Construction

[0030] I. Definition

[0031] In order to understand the density of the present invention, the following terms are defined.

[0032] The terms "bi-biotin", "bi-biotin tag", "bi-B tag" and "bi-biotin moiety" are used interchangeably and generally refer to attachment (optionally covalent attachment) to a reactant of interest of 2 covalently linked biotins. In certain preferred embodiments, the reactant of interest comprises a sequence recognized by a biotin ligase, which catalyzes the covalent attachment of the sequence to a biotin molecule. This sequence is generally referred to as the biotin ligase recognition sequence. Each biotin ligase recognition sequence in a reactant of interest can be covalently linked to a biotin moiety, thus a reactant with multiple biotin ligase recognition sequences can be covalently linked to multiple biotins. A region of a reactant having one or more biotin ligase recognition sequences generally refers to the biotinylated region of the reacta...

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Abstract

Multi-biotinylated reactants are provided which can be used in divalent complexes for various applications such as colocalization, labeling, immobilization, and purification. Methods for constructing, purifying, and using the bis-biotinylated reactants are also provided. In certain embodiments, two bis-biotinylated reactants are bound to a single streptavidin tetramer to provide a complex having a 1:1 stoichiometry with respect to the bis-biotinylated reactants.

Description

[0001] Cross References to Related Applications [0002] This application claims the benefit of US Provisional Application No. 61 / 835,311, filed June 14, 2013, which is hereby incorporated by reference in its entirety for all purposes. [0003] Statement Regarding Federally Funded Research [0004] Not applied. Background technique [0005] A common task in molecular biology is to identify and quantify the presence of proteins in complex mixtures. For example, to identify the expression level of a protein of interest, a Western blot can be performed in which protein extracts are run on a gel and stained with antibodies directed against defined epitopes of the protein of interest. A defined epitope may be a specific sequence or structure found in a native protein, or it may be a tag introduced during cloning, such as a "FLAG tag", against which antibodies are commercially available. A peroxidase-conjugated secondary antibody specific for the primary antibody bound to the pro...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00G01N33/68
CPCG01N33/58C07K14/36
Inventor E·米勒S·卡姆特卡K·比约森J·哈尼斯
Owner PACIFIC BIOSCIENCES
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