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    Quantitative control of sialylation and specific mono-sialylation
    2.
    发明授权
    Quantitative control of sialylation and specific mono-sialylation 有权
    唾液酸化和特异性单唾液酸化的定量控制

    公开(公告)号：US09481902B2

    公开(公告)日：2016-11-01

    申请号：US14970734

    申请日：2015-12-16

    Applicant: Roche Diagnostics Operations, Inc.

    Inventor： Tibor Czabany ,  Alfred Engel ,  Michael Greif ,  Christine Jung ,  Christiane Luley ,  Sebastian Malik ,  Rainer Mueller ,  Bernd Nidetzky ,  Doris Ribitsch ,  Katharina Schmoelzer ,  Helmut Schwab ,  Harald Sobek ,  Bernhard Suppmann ,  Marco Thomann ,  Sabine Zitzenbacher

    IPC: C07K16/00 ,  C12Q1/00 ,  C12P21/00 ,  C12N9/10

    CPC classification number: C12P21/005 ,  C07K16/00 ,  C07K2317/14 ,  C07K2317/24 ,  C07K2317/41 ,  C12N9/1081 ,  C12P19/18 ,  C12Y204/99001

    Abstract: The present disclosure is directed to the use of certain glycosyltransferase variants having N-terminal truncation deletions. It was found that the combination of two different truncation variants of human β-galactoside-α-2,6-sialyltransferase I (hST6Gal-I) exhibited different specific sialyltransferase enzymatic activities. In one example, under conditions wherein the first variant Δ89 hST6Gal-I catalyzed formation of bi-sialylated target molecules the second variant Δ108 hST6Gal-I catalyzed formation of mono-sialylated target molecules. Thus, disclosed are variants of mammalian glycosyltransferase, nucleic acids encoding the same, methods and means for recombinantly producing the variants of mammalian glycosyltransferase and use thereof, particularly for sialylating in a quantitatively controlled manner terminal acceptor groups of glycan moieties being part of glycoproteins such as immunoglobulins.

    Abstract translation: 本公开涉及使用具有N-末端截短缺失的某些糖基转移酶变体。 发现人β-半乳糖苷-α-2,6-唾液酸转移酶I（hST6Gal-1）的两种不同截短变体的组合表现出不同的特异性唾液酸转移酶酶活性。 在一个实例中，在第一变体Δ89hST6Gal-1催化双唾液酸化靶分子的形成的条件下，第二变体Δ108hST6Gal-1催化单唾液酸化靶分子的形成。 因此，公开了哺乳动物糖基转移酶的变体，编码它们的核酸，用于重组产生哺乳动物糖基转移酶变体的方法和手段及其用途，特别是以定量控制的方式唾液酸化作为糖蛋白部分的聚糖部分的末端受体基团，例如 免疫球蛋白。

    QUANTITATIVE CONTROL OF SIALYLATION
    3.
    发明申请
    QUANTITATIVE CONTROL OF SIALYLATION 有权
    SIALYLATION的量化控制

    公开(公告)号：US20160076068A1

    公开(公告)日：2016-03-17

    申请号：US14950443

    申请日：2015-11-24

    Applicant: Roche Diagnostics Operations, Inc.

    Inventor： Alfred Engel ,  Michael Greif ,  Christine Jung ,  Sebastian Malik ,  Rainer Mueller ,  Harald Sobek ,  Bernhard Suppmann ,  Marco Thomann

    IPC: C12P19/18 ,  C12P19/02

    CPC classification number: C12P19/18 ,  C12N9/1081 ,  C12P19/02 ,  C12P19/44 ,  C12P21/005

    Abstract: The present disclosure is directed to the use of certain glycosyltransferase variants having N-terminal truncation deletions. Contrary to previous findings certain truncations were found to exhibit sialidase enzymatic activity, particularly a variant of human sialyltransferase (hST6Gal-I) with a truncation deletion involving the first 89 N-terminal amino acids of the respective wild-type polypeptide. A fundamental finding documented in the present disclosure is that there exists a variant of this enzyme which is capable of catalyzing transfer of a glycosyl moiety as well as hydrolysis thereof. Thus, disclosed is a specific exemplary variant of mammalian glycosyltransferase, nucleic acids encoding the same, methods and means for recombinantly producing the variant of mammalian glycosyltransferase and use thereof, particularly for sialylating in a quantitatively controlled manner terminal acceptor groups of glycan moieties being part of glycoproteins such as immunoglobulins.

    Abstract translation: 本公开涉及使用具有N-末端截短缺失的某些糖基转移酶变体。 发现某些截短表现出唾液酸酶酶活性，特别是具有涉及相应野生型多肽的前89个N-末端氨基酸的截短缺失的人唾液酸转移酶（hST6Gal-1）的变体。 在本公开文献中记载的基本发现是存在能够催化糖基部分的转移以及其水解的酶的变体。 因此，公开了哺乳动物糖基转移酶的特定示例性变体，编码哺乳动物糖基转移酶的核酸，用于重组产生哺乳动物糖基转移酶变体的方法和手段及其用途，特别是以定量控制的方式唾液酸化作为部分的聚糖部分的末端受体基团 糖蛋白如免疫球蛋白。

    THERMOSTABLE AFFINITY POLYPEPTIDES
    4.
    发明公开
    THERMOSTABLE AFFINITY POLYPEPTIDES 审中-公开

    公开(公告)号：US20240140995A1

    公开(公告)日：2024-05-02

    申请号：US18542054

    申请日：2023-12-15

    Applicant: Roche Diagnostics Operations, Inc.

    Inventor： Romina Eisenhauer ,  Alfred Engel ,  Ute Jucknischke ,  Michael Schraeml ,  Wojtek Steffen

    IPC: C07K14/195 ,  G01N33/543

    CPC classification number: C07K14/195 ,  G01N33/54353 ,  C07K2319/00 ,  G01N2333/245

    Abstract: The present invention relates to a method of determining an analyte in a sample, said method comprising (a) contacting said sample with (i) a binding compound binding to said analyte, said binding compound comprising a binding agent and a first partner of an affinity pair (first affinity partner); and (ii) a second partner of the affinity pair (second affinity partner) coupled to a solid surface, to an indicator, and/or to a second binding agent; and (b) determining said analyte based on complexes formed in step (a); wherein one of said first affinity partner and said second affinity partner is a polypeptide comprising the amino acid sequence of SEQ ID NO:1 or a sequence at least 50% identical thereto, and wherein the other of said first affinity partner and said second affinity partner is a polypeptide comprising the amino acid sequence of SEQ ID NO:2 or a sequence at least 50% identical thereto. The present invention also relates to a polypeptide comprising an amino acid sequence as specified in SEQ ID NO:1 or a sequence at least 50% identical thereto, wherein the amino acid at the position corresponding to position 77 in SEQ ID NO:1 is not a histidine; and to a polypeptide comprising an amino acid sequence as specified in SEQ ID NO:2 or a sequence at least 50% identical thereto, wherein (i) the amino acid at the position corresponding to position 17 in SEQ ID NO:2 is not a cysteine, in an embodiment is alanine, serine, leucine, isoleucine, or glycine and/or (ii) said polypeptide further comprises at least one functional peptide; and to fusion polypeptides, polypeptide complexes, polynucleotides and kit related to the aforesaid.

    QUANTITATIVE CONTROL OF SIALYLATION AND SPECIFIC MONO-SIALYLATION
    9.
    发明申请
    QUANTITATIVE CONTROL OF SIALYLATION AND SPECIFIC MONO-SIALYLATION 有权
    SIALYLATION和特定单稳态的量化控制

    公开(公告)号：US20160102333A1

    公开(公告)日：2016-04-14

    申请号：US14970734

    申请日：2015-12-16

    Applicant: Roche Diagnostics Operations, Inc.

    Inventor： Tibor Czabany ,  Alfred Engel ,  Michael Greif ,  Christine Jung ,  Christiane Luley ,  Sebastian Malik ,  Rainer Mueller ,  Bernd Nidetzky ,  Doris Ribitsch ,  Katharina Schmoelzer ,  Helmut Schwab ,  Harald Sobek ,  Bernhard Suppmann ,  Marco Thomann ,  Sabine Zitzenbacher

    IPC: C12P21/00 ,  C07K16/00 ,  C12N9/10

    CPC classification number: C12P21/005 ,  C07K16/00 ,  C07K2317/14 ,  C07K2317/24 ,  C07K2317/41 ,  C12N9/1081 ,  C12P19/18 ,  C12Y204/99001

    Abstract: The present disclosure is directed to the use of certain glycosyltransferase variants having N-terminal truncation deletions. It was found that the combination of two different truncation variants of human β-galactoside-α-2,6-sialyltransferase I (hST6Gal-I) exhibited different specific sialyltransferase enzymatic activities. In one example, under conditions wherein the first variant Δ89 hST6Gal-I catalyzed formation of bi-sialylated target molecules the second variant Δ108 hST6Gal-I catalyzed formation of mono-sialylated target molecules. Thus, disclosed are variants of mammalian glycosyltransferase, nucleic acids encoding the same, methods and means for recombinantly producing the variants of mammalian glycosyltransferase and use thereof, particularly for sialylating in a quantitatively controlled manner terminal acceptor groups of glycan moieties being part of glycoproteins such as immunoglobulins.

    Abstract translation: 本公开涉及使用具有N-末端截短缺失的某些糖基转移酶变体。 发现人和半乳糖苷-α-2,6-唾液酸转移酶I（hST6Gal-1）的两种不同截短变体的组合表现出不同的特异性唾液酸转移酶酶活性。 在一个实例中，在第一变体和Dgr; 89hST6Gal-1催化双唾液酸化靶向分子的形成的条件下，第二变体Dgr; 108hST6Gal-1催化单唾液酸化靶分子的形成。 因此，公开了哺乳动物糖基转移酶的变体，编码它们的核酸，用于重组产生哺乳动物糖基转移酶变体的方法和手段及其用途，特别是以定量控制的方式唾液酸化作为糖蛋白部分的聚糖部分的末端受体基团，例如 免疫球蛋白。

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