Method for detecting and quantifying beta-1,6-branched beta-1,3-glucan or beta-1,3-glucan, and kit for detecting and quantifying the same
AssigneeToei Shinyaku Co., Ltd.
InventorsDaisuke Yamanaka, Yoshiyuki Adachi
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Abstract
In a first step, when a β-1,6-branched β-1,3-glucan is contained in a test specimen, an active reporter protein comprising one and the other of split reporter proteins is formed by binding the β-1,6-branched β-1,3-glucan to a first fusion protein and a second fusion protein, and when a β-1,3-glucan is contained in the test specimen, the active reporter protein comprising one and the other of the split reporter proteins is formed by binding the β-1,3-glucan to the first fusion protein and a third fusion protein, and in a second step, the active reporter protein formed in the first step is detected and quantified.
Claims5
A method for detecting and quantifying a β-1,6-branched β-1,3-glucan or a β-1,3-glucan, the method comprising:
- a first step of bringing a test specimen into contact with a reagent, wherein the reagent contains a reagent (A) and a reagent (B),
- wherein the reagent (A) comprises a first fusion protein, a second fusion protein and a split reporter protein that has a first subunit and other subunit, wherein the first and other subunits form a pair, and the first fusion protein is linked to the first subunit of the split reporter protein and the second fusion protein is linked to the other subunit of the split reporter protein,
- wherein the reagent (B) comprises the first fusion protein, a third fusion protein and the split reporter protein that has a first subunit and other subunit, the first fusion protein is linked to the first subunit of the split reporter protein, the third fusion protein is linked to the other subunit of the split reporter protein,
- wherein the first and other subunits of the split reporter protein form an active reporter protein by making a pair of the first and other subunits of the split reporter protein,
- the first fusion protein comprises a β-1,3-glucan-binding protein,
- the second fusion protein comprises a β-1,6-glucan-binding protein that is a β-1,6-glucanase mutant having no cleavage activity of a β-1,6-glucan and having specific binding activity to a β-1,6-glucan,
- the third fusion protein comprises a β-1,3-glucan-binding protein,
- in the first step, when a β-1,6-branched β-1,3-glucan is contained in the test specimen, the active reporter protein comprising the first subunit and the other subunit of the split reporter protein pair is formed as a result of binding of the β-1,6-branched β-1,3-glucan to the first fusion protein and the second fusion protein of reagent (A),
- when a β-1,3-glucan is contained in the test specimen, the active reporter protein comprising the first subunit and the other subunit of the split reporter protein pair is formed as a result of binding of the β-1,3-glucan to the first fusion protein and the third fusion protein of reagent (B);
- and
- a second step of detecting and quantifying a product obtained by performing the first, step wherein the product comprises a detectable signal produced by the formation of the active reporter protein.
A kit for detecting and quantifying a β-1,6-branched β-1,3-glucan or a β-1,3-glucan, the kit comprising a reagent (A) and a reagent (B),
- wherein the reagent (A) comprises a first fusion protein, a second fusion protein and a split reporter protein that has a first subunit and other subunit, wherein the first and other subunits form a pair, and the first fusion protein is linked to the first subunit of the split reporter protein and the second fusion protein is linked to the other subunit of the split reporter protein,
- wherein the reagent (B) comprises the first fusion protein, a third fusion protein and the split reporter protein that has a first subunit and other subunit, the first fusion protein is linked to the first subunit of the split reporter protein, the third fusion protein is linked to the other subunit of the split reporter protein,
- wherein the first and other subunits of the split reporter protein form an active reporter protein by making a pair of the first and other subunits of the split reporter protein,
- the first fusion protein comprises a β-1,3-glucan-binding protein,
- the second fusion protein comprises a β-1,6-glucan-binding protein that is a β-1,6-glucanase mutant having no cleavage activity of a β-1,6-glucan and having specific binding activity to a β-1,6-glucan,
- the third fusion protein comprises a β-1,3-glucan-binding protein.
Dependencies are inferred from the claim text.