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公开(公告)号：US20160282318A1
公开(公告)日：2016-09-29
申请号：US15066429
申请日：2016-03-10
Applicant: GE Healthcare Bio-Sciences AB
Inventor： Gunnar Malmquist , Martin Hall , Nils Stafstrom
Abstract: The invention discloses a method for correcting a baseline in a chromatogram obtained using a buffer with at least one UV-absorbing component which is a weak acid or a weak base. The method comprises the steps of:a) providing a chromatography system with a chromatography column and a UV detector downstream of the chromatography column; b) providing UV extinction coefficients at a wavelength λ for the acid form and the base form of the UV-absorbing component(s) and a dissociation constant for the component(s), c) obtaining a chromatogram with the UV detector at wavelength λ using the buffer under a set of conditions where pH and/or conductivity is varied between these conditions, d) for a plurality of said conditions calculating the concentrations of the acid and base forms using the dissociation constant(s) and values for pH and conductivity values, and e) calculating UV absorption values of the acid and base forms from the concentrations and UV extinction coefficients under the conditions in question and subtracting the UV absorption values from the chromatogram.
Abstract translation: 一种用于使用具有至少一种UV吸收组分（其为弱酸或弱碱）的缓冲液获得的色谱图中校正基线的方法，包括以下步骤：a）提供具有色谱柱和UV检测器的色谱系统 色谱柱下游; b）为UV形吸收组分的酸形式和碱性形式提供波长为λ的紫外消光系数和组分的解离常数; c）使用UV检测器在波长λ下使用缓冲液在这些条件下pH和/或电导率变化的一组条件下获得色谱图; d）对于多个所述条件，使用解离常数和pH和电导率值计算酸和碱形式的浓度，以及e）从浓度和UV计算酸和碱形式的UV吸收值 在所述条件下的消光系数，并从色谱图中减去紫外吸收值。 酸和碱形式的浓度可以使用Debye-Huckel方程计算。 还要求的是用于执行如上所述的方法的色谱系统。
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公开(公告)号：US10598640B2
公开(公告)日：2020-03-24
申请号：US15066429
申请日：2016-03-10
Applicant: GE Healthcare Bio-Sciences AB
Inventor： Gunnar Malmquist , Martin Hall , Nils Stafstrom
Abstract: The invention discloses a method for correcting a baseline in a chromatogram obtained using a buffer with at least one UV-absorbing component which is a weak acid or a weak base. The method comprises the steps of:a) providing a chromatography system with a chromatography column and a UV detector downstream of the chromatography column;b) providing UV extinction coefficients at a wavelength λ for the acid form and the base form of the UV-absorbing component(s) and a dissociation constant for the component(s),c) obtaining a chromatogram with the UV detector at wavelength λ using the buffer under a set of conditions where pH and/or conductivity is varied between these conditions,d) for a plurality of said conditions calculating the concentrations of the acid and base forms using the dissociation constant(s) and values for pH and conductivity values, ande) calculating UV absorption values of the acid and base forms from the concentrations and UV extinction coefficients under the conditions in question and subtracting the UV absorption values from the chromatogram.
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3.发明申请Chromatography Ligand Comprising Domain C From Staphylococcus Aureus Protein A For Antibody Isolation 有权色谱配体包含来自金黄色葡萄球菌蛋白A的抗体分离区域C公开(公告)号：US20160200797A1
公开(公告)日：2016-07-14
申请号：US15063471
申请日：2016-03-07
Applicant: GE HEALTHCARE BIO-SCIENCES AB
Inventor： Martin Hall , Sture Larsson , Andreas Muranyi , Gustav Rodrigo , Jinyu Zou , Per-Mikael Aberg
CPC classification number: B01J20/24 , B01D15/3809 , B01J20/28019 , B01J20/285 , B01J20/286 , B01J20/289 , B01J20/3212 , B01J20/3219 , B01J20/3274 , B01J20/3293 , B01J2220/52 , B01J2220/54 , C07K1/22 , C07K14/31 , C07K16/00 , C07K17/00 , C07K17/10 , C07K2317/55
Abstract: The present invention relates to a chromatography ligand, which comprises Domain C from Staphylococcus protein A (SpA), or a functional fragment or variant thereof. The chromatography ligand presents an advantageous capability of withstanding harsh cleaning in place (CIP) conditions, and is capable of binding Fab fragments of antibodies. The ligand may be provided with a terminal coupling group, such as arginine or cysteine, to facilitate its coupling to an insoluble carrier such as beads or a membrane. The invention also relates to a process of using the ligand in isolation of antibodies, and to a purification protocol which may include washing steps and/or regeneration with alkali.
Abstract translation: 本发明涉及一种色谱配体，其包含来自葡萄球菌蛋白A（SpA）的结构域C，或其功能片段或变体。 色谱配体具有在现场（CIP）条件下耐受苛刻清洁的有利能力，并且能够结合抗体的Fab片段。 配体可以具有末端偶联基团，例如精氨酸或半胱氨酸，以促进其与不溶性载体例如珠粒或膜的偶联。 本发明还涉及在分离抗体中使用配体的方法以及可以包括洗涤步骤和/或用碱再生的纯化方案。
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4.发明授权Chromatography ligand comprising domain C from Staphylococcus aureus protein A for antibody isolation 有权包含来自金黄色葡萄球菌蛋白A的结构域C的色谱配体用于抗体分离公开(公告)号：US09290549B2
公开(公告)日：2016-03-22
申请号：US14164519
申请日：2014-01-27
Applicant: GE HEALTHCARE BIO-SCIENCES AB
Inventor： Martin Hall , Sture Larsson , Andreas Muranyi , Gustav Rodrigo , Jinyu Zou , Per-Mikael Aberg
CPC classification number: B01J20/24 , B01D15/3809 , B01J20/28019 , B01J20/285 , B01J20/286 , B01J20/289 , B01J20/3212 , B01J20/3219 , B01J20/3274 , B01J20/3293 , B01J2220/52 , B01J2220/54 , C07K1/22 , C07K14/31 , C07K16/00 , C07K17/00 , C07K17/10 , C07K2317/55
Abstract: The present invention relates to a chromatography ligand, which comprises Domain C from Staphylococcus protein A (SpA), or a functional fragment or variant thereof. The chromatography ligand presents an advantageous capability of withstanding harsh cleaning in place (CIP) conditions, and is capable of binding Fab fragments of antibodies. The ligand may be provided with a terminal coupling group, such as arginine or cysteine, to facilitate its coupling to an insoluble carrier such as beads or a membrane. The invention also relates to a process of using the ligand in isolation of antibodies, and to a purification protocol which may include washing steps and/or regeneration with alkali.
Abstract translation: 本发明涉及一种色谱配体，其包含来自葡萄球菌蛋白A（SpA）的结构域C，或其功能片段或变体。 色谱配体具有在现场（CIP）条件下耐受苛刻清洁的有利能力，并且能够结合抗体的Fab片段。 配体可以具有末端偶联基团，例如精氨酸或半胱氨酸，以促进其与不溶性载体例如珠粒或膜的偶联。 本发明还涉及在分离抗体中使用配体的方法以及可以包括洗涤步骤和/或用碱再生的纯化方案。
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