JPWO2019185515A5 - - Google Patents

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JPWO2019185515A5
JPWO2019185515A5 JP2021501084A JP2021501084A JPWO2019185515A5 JP WO2019185515 A5 JPWO2019185515 A5 JP WO2019185515A5 JP 2021501084 A JP2021501084 A JP 2021501084A JP 2021501084 A JP2021501084 A JP 2021501084A JP WO2019185515 A5 JPWO2019185515 A5 JP WO2019185515A5
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antibody
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biomarker protein
fucα1
cancer
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タンパク質の糖鎖プロファイルを判定する方法であって、
(a)前記タンパク質を含むサンプルを、前記タンパク質に対する抗体に接触させて、抗体-タンパク質複合体を形成するステップと、
(b)ステップ(a)で得た前記抗体-タンパク質複合体を単離するステップと、
(c)その後、前記抗体-タンパク質複合体を1つ以上のレクチンに接触させて、前記タンパク質の糖鎖プロファイルを判定するステップとを含み、
ステップ(a)の前記抗体は固体表面上に固定化されておらず、前記タンパク質は、該方法を行っているとき、前記抗体から放出されない、方法。
A method for determining the sugar chain profile of a protein.
(A) A step of contacting a sample containing the protein with an antibody against the protein to form an antibody-protein complex.
(B) The step of isolating the antibody-protein complex obtained in step (a) and
(C) The step comprising contacting the antibody-protein complex with one or more lectins to determine the sugar chain profile of the protein.
The method, wherein the antibody of step (a) is not immobilized on a solid surface and the protein is not released from the antibody when performing the method.
(d)前記タンパク質の糖鎖プロファイルを前記タンパク質の対照糖鎖プロファイルと比較して、前記タンパク質の糖鎖プロファイルが前記対照糖鎖プロファイルの糖鎖プロファイルからずれ得るかどうかを判定するステップをさらに含む、請求項1に記載の方法。 (D) Further comprising a step of comparing the sugar chain profile of the protein with the control sugar chain profile of the protein to determine whether the sugar chain profile of the protein can deviate from the sugar chain profile of the control sugar chain profile. , The method according to claim 1. 前記タンパク質は、がんバイオマーカータンパク質、自己免疫性疾患バイオマーカータンパク質、又は炎症性疾患バイオマーカータンパク質である、請求項1又は2に記載の方法。 The method according to claim 1 or 2, wherein the protein is a cancer biomarker protein, an autoimmune disease biomarker protein, or an inflammatory disease biomarker protein. 前記がんバイオマーカータンパク質は、卵巣がんバイオマーカータンパク質、乳がんバイオマーカータンパク質、結腸直腸がんバイオマーカータンパク質、膵がんバイオマーカータンパク質、前立腺がんバイオマーカータンパク質、甲状腺がんバイオマーカータンパク質、肝がんバイオマーカータンパク質、肺がんバイオマーカータンパク質、胃がんバイオマーカータンパク質、精巣がんバイオマーカータンパク質、又は膀胱がんバイオマーカータンパク質である、請求項3に記載の方法。 The cancer biomarker proteins include ovarian cancer biomarker protein, breast cancer biomarker protein, colon-rectal cancer biomarker protein, pancreatic cancer biomarker protein, prostate cancer biomarker protein, thyroid cancer biomarker protein, and liver. The method according to claim 3, which is a cancer biomarker protein, a lung cancer biomarker protein, a stomach cancer biomarker protein, a testis cancer biomarker protein, or a bladder cancer biomarker protein. 前記前立腺がんバイオマーカータンパク質はβ-ハプトグロビン、TIMP-1、PSA、fPSA、又はtPSAである、請求項4に記載の方法。 The method of claim 4, wherein the prostate cancer biomarker protein is β-haptoglobin, TIMP-1, PSA, fPSA, or tPSA. 前記抗体は、前記抗体の単離を可能にするビーズを含む、請求項1~5のいずれか1項に記載の方法。 The method according to any one of claims 1 to 5, wherein the antibody comprises beads that enable isolation of the antibody. 前記1つ以上のレクチンは、コアフコース、アンテナ型フコース、Fucα1-6GlcNAc-N-Asn含有N-結合型オリゴ糖、Fucα1-6/3GlcNAc、α-L-Fuc、Fucα1-2Galβ1-4(Fucα1-3)GlcNAc、Fucα1-2Gal、Fucα1-6GlcNAc、Manβ1-4GlcNAcβ1-4GlcNAc、分岐N-結合型6糖、Manα1-3Man、α-D-Man、(GlcNAcβ1-4)2-4、Galβ1-4GlcNAc、GlcNAcα1-4Galβ1-4GlcNAc、(GlcNAcβ1-4)2-5、Neu5Ac(シアル酸)、Galβ1-3GalNAc-セリン/スレオニン、Galα1-3GalNAc、Galβ1-6Gal、Galβ1-4GlcNAc、Galβ1-3GalNAc、GalNAcα1-3GalNAc、GalNAcα1-3Gal、GalNAcα/β1-3/4Gal、α-GalNAc、GalNAcβ1-4Gal、GalNAcα1-3(Fucα1-2)Gal、GalNAcα1-2Gal、GalNAcα1-3GalNAc、GalNAcβ1-3/4Gal、GalNAc-Ser/Thr (Tn抗原)、Galβ1-3GalNAc-Ser/Thr (T抗原)、GalNAcβ1-4GlcNAc (LacdiNAc)、α-2,3Neu5Ac(α2-3結合シアル酸)、α-2,6Neu5Ac(α2-6結合シアル酸)、α-2,8Neu5Ac(α2-8結合シアル酸)、シアル酸(α-2,3Neu5Ac、α-2,6Neu5Ac又はα-2,8Neu5Ac)、Neu5Acα4/9-O-Ac-Neu5Ac、Neu5Acα2-3Galβ1-4Glc/GlcNAc、Neu5Acα2-6Gal/GalNAc、N-結合2アンテナ型、N-結合3/4アンテナ型、分岐β1-6GlcNAc、Galα1-3(Fucα1-2)Galβ1-3/4GlcNAc、Galβ1-3(Fucα1-4)GlcNAc、NeuAcα2-3Galβ1-3(Fucα1-4)GlcNAc、Fucα1-2Galβ1-3(Fucα1-4)GlcNAc、Galβ1-4(Fucα1-3)GlcNAc、NeuAcα2-3Galβ1-4(Fucα1-3)GlcNAc、Fucα1-2Galβ1-4(Fucα1-3)GlcNAc、高マンノース、シアリルルイスa(シアリルLea)抗原、シアリルルイスx(シアリルLex)抗原、ルイスx(Lex) 抗原、シアリルTn抗原、シアリルT抗原、ルイスy(Ley)抗原、硫酸化コア1糖鎖、Tn抗原、T抗原、コア2糖鎖、ルイスa(Lea) 抗原、(GlcNAcβ1-4)n、β-D-GlcNAc、GalNAc、Gal-GlcNAc、GlcNAc、Galα1-3Gal、Galβ1-3GalNAc、α-Gal、α-GalNAc、(GlcNAc)n、分岐(LacNAc)nに特異的である、請求項1~6のいずれか1項に記載の方法。 The one or more lectins are core fucose, antenna type fucose, Fucα1-6GlcNAc-N-Asn-containing N-linked oligosaccharide, Fucα1-6 / 3GlcNAc, α-L-Fuc, Fucα1-2Galβ1-4 (Fucα1-3). ) GlcNAc, Fucα1-2Gal, Fucα1-6GlcNAc, Manβ1-4GlcNAcβ1-4GlcNAc, branched N-linked 6 sugars, Manα1-3Man, α-D-Man, (GlcNAcβ1-4) 2-4, Galβ1-4GlcNAc, GlcNAcα1- 4Galβ1-4GlcNAc, (GlcNAcβ1-4) 2-5, Neu5Ac (sialic acid), Galβ1-3GalNAc-serine / threonine, Galα1-3GalNAc, Galβ1-6Gal, Galβ1-4GlcNAc, Galβ1-3GalNAc, GalNAcα1-3Gal , GalNAcα / β1-3 / 4Gal, α-GalNAc, GalNAcβ1-4Gal, GalNAcα1-3 (Fucα1-2) Gal, GalNAcα1-2Gal, GalNAcα1-3GalNAc, GalNAcβ1-3 / 4Gal, GalNAc-Ser / Thr (Tn antigen) , Galβ1-3GalNAc-Ser / Thr (T antigen), GalNAcβ1-4GlcNAc (LacdiNAc), α-2,3Neu5Ac (α2-3 bound sialic acid), α-2,6Neu5Ac (α2-6 bound sialic acid), α- 2,8Neu5Ac (α2-8 bound sialic acid), sialic acid (α-2,3Neu5Ac, α-2,6Neu5Ac or α-2,8Neu5Ac), Neu5Ac α4 / 9-O-Ac-Neu5Ac, Neu5Ac α2-3Gal β1-4Glc / GlcNAc, Neu5Acα2-6Gal / GalNAc, N-coupled 2 antenna type, N-coupled 3/4 antenna type, branched β1-6GlcNAc, Galα1-3 (Fucα1-2) Galβ1-3 / 4GlcNAc, Galβ1-3 (Fucα1-4) ) GlcNAc, NeuAcα2-3Galβ1-3 (Fucα1-4) GlcNAc, Fucα1-2Galβ1-3 (Fucα1-4) GlcNAc, Galβ1-4 (Fucα1-3) GlcNAc, NeuAcα2-3Galβ1-4 (Fucα1-3) GlcNAc, Fucα1 -2Galβ1-4 (Fucα1-3) GlcNAc, high Mannose, sialyl Lewis a (sialyl Lea) antigen, sialyl Lewis x (sialyl Lex) antigen, Lewis x (Lex) antigen, sialyl Tn antigen, sialyl T antigen, Lewis y (Ley) antigen, sulfated core 1 sugar chain, Tn antigen, T antigen, core 2 sugar chain, Lewis a (Lea) antigen, (GlcNAcβ1-4) n, β-D-GlcNAc, GalNAc, Gal-GlcNAc, GlcNAc, Galα1-3Gal, Galβ1-3GalNAc, α-Gal, α- The method according to any one of claims 1 to 6, which is specific for GalNAc, (GlcNAc) n, and branch (LacNAc) n. 対象ががんのリスクを有し得るか、又はがんに罹患し得ることを診断する方法であって、
(a)前記対象から得た、がんバイオマーカータンパク質を含むサンプルを、前記がんバイオマーカータンパク質に対する抗体に接触させて、抗体-がんバイオマーカータンパク質複合体を形成するステップと、
(b)ステップ(a)で得た前記抗体-タンパク質複合体を単離するステップと、
(c)その後、前記抗体-がんバイオマーカータンパク質複合体を1つ以上のレクチンに接触させて、前記がんバイオマーカータンパク質の糖鎖プロファイルを判定するステップとを含み、
ステップ(a)の前記抗体は固体表面上に固定化されておらず、前記タンパク質は、該方法を行っているとき、前記抗体から放出されず、
前記糖鎖プロファイルの、前記がんバイオマーカータンパク質の健常な糖鎖プロファイルからのずれは、前記対象ががんのリスクを有し得る、又はがんに罹患し得ることを示す、方法。
A method of diagnosing that a subject may be at risk of cancer or may have cancer.
(A) A step of contacting a sample containing a cancer biomarker protein obtained from the subject with an antibody against the cancer biomarker protein to form an antibody-cancer biomarker protein complex.
(B) The step of isolating the antibody-protein complex obtained in step (a) and
(C) The step comprising contacting the antibody-cancer biomarker protein complex with one or more lectins to determine the sugar chain profile of the cancer biomarker protein.
The antibody of step (a) was not immobilized on a solid surface and the protein was not released from the antibody when performing the method.
A method of indicating that a deviation of the sugar chain profile from a healthy sugar chain profile of the cancer biomarker protein indicates that the subject may be at risk of cancer or may suffer from cancer.
対象が自己免疫性疾患のリスクを有し得るか、又は自己免疫性疾患に罹患し得ることを診断する方法であって、
(a)前記対象から得た、自己免疫性疾患バイオマーカータンパク質を含むサンプルを、前記自己免疫性疾患バイオマーカータンパク質に対する抗体に接触させて、抗体-自己免疫性疾患バイオマーカータンパク質複合体を形成するステップと、
(b)ステップ(a)で得た前記抗体-タンパク質複合体を単離するステップと、
(c)その後、前記抗体-自己免疫性疾患バイオマーカータンパク質複合体を1つ以上のレクチンに接触させて、前記自己免疫性疾患バイオマーカータンパク質の糖鎖プロファイルを判定するステップとを含み、
ステップ(a)の前記抗体は固体表面上に固定化されておらず、前記タンパク質は、該方法を行っているとき、前記抗体から放出されず、
前記糖鎖プロファイルの、前記自己免疫性疾患バイオマーカータンパク質の健常な糖鎖プロファイルからのずれは、前記対象が自己免疫性疾患のリスクを有し得る、又は自己免疫性疾患に罹患し得ることを示す、方法。
A method of diagnosing that a subject may be at risk for an autoimmune disease or may suffer from an autoimmune disease.
(A) A sample containing an autoimmune disease biomarker protein obtained from the subject is contacted with an antibody against the autoimmune disease biomarker protein to form an antibody-autoimmune disease biomarker protein complex. Steps and
(B) The step of isolating the antibody-protein complex obtained in step (a) and
(C) The step comprising contacting the antibody-autoimmune disease biomarker protein complex with one or more lectins to determine the sugar chain profile of the autoimmune disease biomarker protein.
The antibody of step (a) was not immobilized on a solid surface and the protein was not released from the antibody when performing the method.
Deviations of the glycan profile from the healthy glycan profile of the autoimmune disease biomarker protein indicate that the subject may be at risk for autoimmune disease or may suffer from autoimmune disease. Show, how.
対象が炎症性疾患のリスクを有し得るか、又は炎症性疾患に罹患し得ることを診断する方法であって、
(a)前記対象から得た、炎症性疾患バイオマーカータンパク質を含むサンプルを、前記炎症性疾患バイオマーカータンパク質に対する抗体に接触させて、抗体-炎症性疾患バイオマーカータンパク質複合体を形成するステップと、
(b)ステップ(a)で得た前記抗体-タンパク質複合体を単離するステップと、
(c)その後、前記抗体-炎症性疾患バイオマーカータンパク質複合体を1つ以上のレクチンに接触させて、前記炎症性疾患バイオマーカータンパク質の糖鎖プロファイルを判定するステップとを含み、
ステップ(a)の前記抗体は固体表面上に固定化されておらず、前記タンパク質は、該方法を行っているとき、前記抗体から放出されず、
前記糖鎖プロファイルの、前記炎症性疾患バイオマーカータンパク質の健常な糖鎖プロファイルからのずれは、前記対象が炎症性疾患のリスクを有し得る、又は炎症性疾患に罹患し得ることを示す、方法。
A method of diagnosing that a subject may be at risk of or suffering from an inflammatory disease.
(A) A step of contacting a sample containing the inflammatory disease biomarker protein obtained from the subject with an antibody against the inflammatory disease biomarker protein to form an antibody-inflammatory disease biomarker protein complex.
(B) The step of isolating the antibody-protein complex obtained in step (a) and
(C) The step comprising contacting the antibody-inflammatory disease biomarker protein complex with one or more lectins to determine the sugar chain profile of the inflammatory disease biomarker protein.
The antibody of step (a) was not immobilized on a solid surface and the protein was not released from the antibody when performing the method.
A method of indicating that a deviation of the glycan profile from a healthy glycan profile of the inflammatory disease biomarker protein indicates that the subject may be at risk of inflammatory disease or may suffer from inflammatory disease. ..
請求項4に記載されるがんバイオマーカータンパク質に特異的な抗体と、請求項7に記載される1つ以上のレクチンとを含む、請求項8に記載の方法を行うためのキット。 A kit for performing the method of claim 8, comprising the antibody specific for the cancer biomarker protein of claim 4 and one or more lectins of claim 7. IgGである自己免疫性疾患バイオマーカータンパク質に特異的である抗体と、請求項7に記載される1つ以上のレクチンとを含む、請求項9に記載の方法を行うためのキット。 A kit for performing the method of claim 9, comprising an antibody specific for an autoimmune disease biomarker protein that is IgG and one or more lectins according to claim 7. IgG、IgA、又はCRPである炎症性疾患バイオマーカータンパク質に特異的である抗体と、請求項7に記載される1つ以上のレクチンとを含む、請求項10に記載の方法を行うためのキット。 A kit for performing the method of claim 10, comprising an antibody specific for an inflammatory disease biomarker protein, IgG, IgA, or CRP, and one or more lectins according to claim 7. .. 前記1つ以上のレクチンは、固定化されている、請求項1~10のいずれか1項に記載の方法。The method according to any one of claims 1 to 10, wherein the one or more lectins are immobilized. 前記1つ以上のレクチンは、サンプル位置に固定化されている、請求項14に記載の方法。14. The method of claim 14, wherein the one or more lectins are immobilized at sample positions. 前記レクチンは、磁性担体に結合されている、請求項1~10及び14のいずれか1項に記載の方法。The method according to any one of claims 1 to 10 and 14, wherein the lectin is bound to a magnetic carrier. 前記抗体は、例えばビオチンであるリガンド基で修飾されている、請求項1~10及び14~16のいずれか1項に記載の方法。The method according to any one of claims 1 to 10 and 14 to 16, wherein the antibody is modified with a ligand group, for example, biotin. 前記抗体は、ストレプトアビジンによって検出可能である、請求項1~10及び14~17のいずれか1項に記載の方法。The method according to any one of claims 1 to 10 and 14 to 17, wherein the antibody can be detected by streptavidin. 前記抗体は、磁性担体に結合されている、請求項1~10及び14~18のいずれか1項に記載の方法。The method according to any one of claims 1 to 10 and 14 to 18, wherein the antibody is bound to a magnetic carrier. 前記抗体は、磁性担体に固定化された抗糖タンパク質抗体又はその抗原結合タンパク質であり、The antibody is an anti-glycoprotein antibody immobilized on a magnetic carrier or an antigen-binding protein thereof.
前記磁性担体は、ペルオキシダーゼ活性を有するポリペプチドを含む、請求項1~10及び14~19のいずれか1項に記載の方法。The method according to any one of claims 1 to 10 and 14 to 19, wherein the magnetic carrier contains a polypeptide having peroxidase activity.
前記抗体は、フルオロフォア、酵素、放射性同位体、蛍光タンパク質、蛍光色素、又はタグを含む、請求項1~10及び14~20のいずれか1項に記載の方法。The method according to any one of claims 1 to 10 and 14 to 20, wherein the antibody comprises a fluorophore, an enzyme, a radioisotope, a fluorescent protein, a fluorescent dye, or a tag.
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