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    • 2. 发明申请
    • METHOD FOR DETERMINING THE IN VIVO INTERACTION MODE
    • 确定体内交互模式的方法
    • WO2017097918A1
    • 2017-06-15
    • PCT/EP2016/080292
    • 2016-12-08
    • F. HOFFMANN-LA ROCHE AGHOFFMANN-LA ROCHE INC.
    • MOELLEKEN, JoergMOLHOJ, MichaelGASSNER, ChristianENDESFELDER, ManuelREGULA, Thomas
    • G01N33/569G01N33/53G01N33/536
    • G01N33/53G01N33/536
    • Herein is reported a method for determining the binding interaction with a multimeric antigen of an antibody of the human IgG1 subclass that has at least two binding sites specifically binding to the antigen comprising the steps of 1) determining the binding affinity of the antibody for the multimeric antigen, and 2) incubating a mixture comprising the antibody and a polypeptide that is derived from lysine-gingipain of porphyromonas gingivalis under conditions and for a time sufficient to cleave the antibody into Fabs and Fc-region, and determining the binding affinity of the Fabs of the antibody for the multimeric, whereby the binding affinity of the antibody to the multimeric antigen to be affinity-driven if the binding affinity determined in both steps are comparable and to be avidity-driven if the binding affinity determined in both steps are different.
    • 本文报道了用于确定与具有至少两个特异性结合抗原的结合位点的人IgG1亚类抗体的多聚体抗原的结合相互作用的方法,其包括以下步骤:1)测定 抗体对多聚体抗原的结合亲和力,和2)在足以将抗体裂解成Fab和Fc区的条件下孵育包含抗体和源自牙龈卟啉单胞菌的赖氨酸 - 牙龈卟啉菌素的多肽的混合物, 并确定抗体的Fab对多聚体的结合亲和力,由此如果在两个步骤中确定的结合亲和力是可比较的,则抗体对多聚体抗原的结合亲和力为亲和力驱动的,并且如果结合 在两个步骤中确定的亲和力都是不同的。
    • 3. 发明申请
    • DIRECT AFFINITY MEASUREMENT OF HUMAN IGG1 BINDING MULTIMERIC ANTIGENS
    • 人IGG1结合多重抗原的直接亲和力测定
    • WO2017097706A1
    • 2017-06-15
    • PCT/EP2016/079756
    • 2016-12-05
    • F. HOFFMANN-LA ROCHE AGHOFFMANN-LA ROCHE INC.
    • MOLHOJ, MichaelGASSNER, ChristianMOELLEKEN, JoergENDESFELDER, Manuel
    • G01N33/569G01N33/53G01N33/536
    • G01N33/56955G01N33/53G01N33/536
    • Herein is reported a method for determining the binding affinity of the binding sites of a bivalent full length antibody of the human IgG1 subclass to a homo-multimeric antigen comprising the steps of i) incubating a mixture comprising the antibody and a polypeptide that is derived from lysine-gingipain of porphyromonas gingivalis at a pH of from pH 7.5 to pH 8.5, in the presence of a reducing agent, at a temperature of from 30 °C to 42°C, for time of from 10 min. to 240 min. to cleave the antibody into Fabs and Fc-region, and ii) determining the binding affinity of the Fabs of the antibody for its antigen using a surface plasmon resonance method by directly applying the incubated reaction mixture obtained in the previous step in the surface plasmon resonance method and therewith determining the binding affinity of the binding sites of the bivalent full length antibody of the human IgG1 subclass.
    • 本文报道了测定人IgG1亚类的二价全长抗体的结合位点与同源多聚体抗原的结合亲和力的方法,其包括以下步骤:i)将包含 抗体和多肽,其在pH为7.5至pH 8.5的pH下,在还原剂存在下,在30℃至42℃的温度下,从牙龈卟啉单胞菌的赖氨酸 - 牙龈卟啉菌衍生,持续时间为 从10分钟。 到240分钟。 将抗体切割成Fab和Fc区,以及ii)使用表面等离子体共振方法通过直接将前一步骤中获得的温育的反应混合物应用于表面等离子体共振来确定抗体的Fab对其抗原的结合亲和力 方法,并由此确定人IgG1亚类的二价全长抗体的结合位点的结合亲和力。