CN105358228A - 亲和层析基质 - Google Patents
亲和层析基质 Download PDFInfo
- Publication number
- CN105358228A CN105358228A CN201480038362.8A CN201480038362A CN105358228A CN 105358228 A CN105358228 A CN 105358228A CN 201480038362 A CN201480038362 A CN 201480038362A CN 105358228 A CN105358228 A CN 105358228A
- Authority
- CN
- China
- Prior art keywords
- oligosaccharides
- blood group
- matrix
- transplanted
- epi
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/06—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies from serum
- C07K16/065—Purification, fragmentation
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
- A61K39/39516—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum from serum, plasma
- A61K39/39525—Purification
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
- B01D15/08—Selective adsorption, e.g. chromatography
- B01D15/26—Selective adsorption, e.g. chromatography characterised by the separation mechanism
- B01D15/38—Selective adsorption, e.g. chromatography characterised by the separation mechanism involving specific interaction not covered by one or more of groups B01D15/265 - B01D15/36
- B01D15/3804—Affinity chromatography
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
- B01D15/08—Selective adsorption, e.g. chromatography
- B01D15/26—Selective adsorption, e.g. chromatography characterised by the separation mechanism
- B01D15/38—Selective adsorption, e.g. chromatography characterised by the separation mechanism involving specific interaction not covered by one or more of groups B01D15/265 - B01D15/36
- B01D15/3804—Affinity chromatography
- B01D15/3809—Affinity chromatography of the antigen-antibody type, e.g. protein A, G, L chromatography
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
- B01J20/22—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof comprising organic material
- B01J20/26—Synthetic macromolecular compounds
- B01J20/265—Synthetic macromolecular compounds modified or post-treated polymers
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
- B01J20/281—Sorbents specially adapted for preparative, analytical or investigative chromatography
- B01J20/286—Phases chemically bonded to a substrate, e.g. to silica or to polymers
- B01J20/289—Phases chemically bonded to a substrate, e.g. to silica or to polymers bonded via a spacer
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
- B01J20/281—Sorbents specially adapted for preparative, analytical or investigative chromatography
- B01J20/291—Gel sorbents
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
- B01J20/30—Processes for preparing, regenerating, or reactivating
- B01J20/32—Impregnating or coating ; Solid sorbent compositions obtained from processes involving impregnating or coating
- B01J20/3214—Impregnating or coating ; Solid sorbent compositions obtained from processes involving impregnating or coating characterised by the method for obtaining this coating or impregnating
- B01J20/3217—Resulting in a chemical bond between the coating or impregnating layer and the carrier, support or substrate, e.g. a covalent bond
- B01J20/3219—Resulting in a chemical bond between the coating or impregnating layer and the carrier, support or substrate, e.g. a covalent bond involving a particular spacer or linking group, e.g. for attaching an active group
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
- B01J20/30—Processes for preparing, regenerating, or reactivating
- B01J20/32—Impregnating or coating ; Solid sorbent compositions obtained from processes involving impregnating or coating
- B01J20/3231—Impregnating or coating ; Solid sorbent compositions obtained from processes involving impregnating or coating characterised by the coating or impregnating layer
- B01J20/3242—Layers with a functional group, e.g. an affinity material, a ligand, a reactant or a complexing group
- B01J20/3244—Non-macromolecular compounds
- B01J20/3246—Non-macromolecular compounds having a well defined chemical structure
- B01J20/3248—Non-macromolecular compounds having a well defined chemical structure the functional group or the linking, spacer or anchoring group as a whole comprising at least one type of heteroatom selected from a nitrogen, oxygen or sulfur, these atoms not being part of the carrier as such
- B01J20/3251—Non-macromolecular compounds having a well defined chemical structure the functional group or the linking, spacer or anchoring group as a whole comprising at least one type of heteroatom selected from a nitrogen, oxygen or sulfur, these atoms not being part of the carrier as such comprising at least two different types of heteroatoms selected from nitrogen, oxygen or sulphur
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
- B01J20/30—Processes for preparing, regenerating, or reactivating
- B01J20/32—Impregnating or coating ; Solid sorbent compositions obtained from processes involving impregnating or coating
- B01J20/3231—Impregnating or coating ; Solid sorbent compositions obtained from processes involving impregnating or coating characterised by the coating or impregnating layer
- B01J20/3242—Layers with a functional group, e.g. an affinity material, a ligand, a reactant or a complexing group
- B01J20/3244—Non-macromolecular compounds
- B01J20/3246—Non-macromolecular compounds having a well defined chemical structure
- B01J20/3248—Non-macromolecular compounds having a well defined chemical structure the functional group or the linking, spacer or anchoring group as a whole comprising at least one type of heteroatom selected from a nitrogen, oxygen or sulfur, these atoms not being part of the carrier as such
- B01J20/3255—Non-macromolecular compounds having a well defined chemical structure the functional group or the linking, spacer or anchoring group as a whole comprising at least one type of heteroatom selected from a nitrogen, oxygen or sulfur, these atoms not being part of the carrier as such comprising a cyclic structure containing at least one of the heteroatoms nitrogen, oxygen or sulfur, e.g. heterocyclic or heteroaromatic structures
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
- B01J20/30—Processes for preparing, regenerating, or reactivating
- B01J20/32—Impregnating or coating ; Solid sorbent compositions obtained from processes involving impregnating or coating
- B01J20/3231—Impregnating or coating ; Solid sorbent compositions obtained from processes involving impregnating or coating characterised by the coating or impregnating layer
- B01J20/3242—Layers with a functional group, e.g. an affinity material, a ligand, a reactant or a complexing group
- B01J20/3268—Macromolecular compounds
- B01J20/3272—Polymers obtained by reactions otherwise than involving only carbon to carbon unsaturated bonds
- B01J20/3274—Proteins, nucleic acids, polysaccharides, antibodies or antigens
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/16—Extraction; Separation; Purification by chromatography
- C07K1/22—Affinity chromatography or related techniques based upon selective absorption processes
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
- C07K16/34—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against blood group antigens
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J2220/00—Aspects relating to sorbent materials
- B01J2220/80—Aspects related to sorbents specially adapted for preparative, analytical or investigative chromatography
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/582—Recycling of unreacted starting or intermediate materials
Abstract
本发明涉及一种呈凝胶形式的亲和层析基质,其包括聚合物颗粒,在这些颗粒上通过间隔基移植了至少一种与A血型和/或B血型表位相对应的寡糖,其特征在于寡糖的密度为0.2至0.7mg/ml的基质。本发明还涉及该基质用于制备具有治疗用途的免疫球蛋白浓缩物的应用。
Description
技术领域
本发明涉及实施亲和层析方法所需的基质,其专门用于阻留及清除血液产品的A血型和/或B血型抗原之抗体。
发明背景
目前,许多疾病均采用免疫球蛋白组合物(Ig)进行治疗。可以举出关于具有抗体生产缺损的原发性免疫缺陷的实例,如:川崎氏病、儿童及成人免疫性血小板减少性紫癜、具有抗体生产缺损的继发性免疫缺陷,尤其为慢性淋巴细胞白血病或与复发性感染相关的骨髓瘤、与细菌感染相关的儿童人类免疫缺陷病毒感染、多灶性运动神经病、格林巴利综合征、严重的急性或慢性人类微小病毒感B19感染、组成性免疫缺陷或获得性免疫缺陷、皮质性皮肌炎、急性重症肌无力症、慢性特发性多神经根神经炎、免疫性血小板减少性紫癜;如与人类免疫缺陷病毒感染相关的疾病,如僵人综合征(Stiffmansyndrome)、自身免疫性嗜中性粒细胞减少症、由自身抗体引起的急性幼红细胞减少症(resistantauto–immuneerythroblastopenia)、自身抗体获得性抗凝症、类风湿性多关节炎、葡萄膜炎等。
在治疗中对富含Ig的人血浆成分的增加的使用需要大量例如从人血浆生产高度纯化的能够进行静脉注射(IgIV)或皮下注射(Igsc)的Ig浓缩物。
该类生产面临着许多制约因素,特别是在所获得Ig浓缩物无害性方面所面临的制约因素。
据欧洲药典记载,为降低溶血危险,应降低这些浓缩物中A血型和/或B血型抗原的抗体(在说明书中简写为“抗A血型抗体”及“抗B血型抗体”或称为“抗-A抗体”及“抗-B抗体”,或称为“抗-A同族凝集素”及“抗-B同族凝集素”)的含量。
由于Ig方面的需求不断增加,有必要设立容量愈来愈大的供体库,其在统计学上将富含O血型。因此,显然有必要需要相关方法,该方法在使用血浆库内血浆来进行Ig浓缩物工业生产的过程中能够消除血液产品中的抗-A抗体及抗-B抗体。
申请WO2007/077365中公开一种Ig浓缩物制备方法,其中包括一种旨在减少抗-A抗体及抗-B抗体的浓缩物的亲和层析步骤。
发明内容
目前,本申请人已开发了一种基质,其尤其有利于在工业化水平上通过亲和层析来阻留及清除血液产品中的抗-A和/或抗-B抗体。
因此,本发明提供一种呈凝胶形式的亲和层析基质,其包括聚合物颗粒;在这些颗粒上移植了至少一种与A血型和/或B血型表位相对应的寡糖;该寡糖通过间隔基移植至该颗粒;其特征在于:寡糖的密度包括在0.2至0.7mg/ml的基质之间。该间隔基的特征在于其具有式(I)-NH-R1-CO-NH-R2;其中,R1为C4-C6烷基,R2为C3-C8烷基;且该间隔基通过其胺官能团与颗粒结合。
这些移植的颗粒的示意图已在图1中给出。
在特定实施方案中,基质包括(i)聚合物颗粒,其上移植了至少一种与A血型表位相对应的寡糖;和/或(ii)聚合物颗粒;其上移植了至少一种与B血型表位相对应的寡糖。
在优选实施方案中,基质包括(i)聚合物颗粒,其上移植了至少一种与A血型表位相对应的寡糖;以及(ii)聚合物颗粒;其上移植了至少一种与B血型表位相对应的寡糖。该实施方案已于图4A中予以阐述。
在另一个实施方案中,基质包括聚合物颗粒;其上移植了至少一种与A血型表位相对应的寡糖,和至少一种与B血型表位相对应的寡糖。该实施方案已于图4B中予以阐述。
在特定实施方案中,基质包括由如下物质组成的混合物:(i)聚合物颗粒,其上移植了至少一种与A血型表位相对应的寡糖;(ii)聚合物颗粒,其上移植了至少一种与B血型表位相对应的寡糖;以及(iii)聚合物颗粒,其上既移植了至少一种与A血型表位相对应的寡糖,也移植至少一种与B血型表位相对应的寡糖。该实施方案已于图4C中予以阐述。
在所有实施方案中,用于将与A血型表位相对应的配体与颗粒连接的间隔基,以及用于将与B血型表位相对应的配体与颗粒连接的间隔基可为相同或不同的,优选为相同的间隔基,但通常选用式(I)中的间隔基。
本发明还涉及该基质或该介质在结合抗-A抗体和/或抗-B抗体的亲和层析方法中的应用。
因此,本发明还旨在该基质或该介质进行具有治疗用途的免疫球蛋白G(IgG)之工业化生产的应用。
本发明还提供一种制备具有治疗用途的免疫球蛋白(Ig)浓缩物的方法,其中包括使用乙醇分级和/或辛酸分级的方法和/或层析分离法自血浆中获得Ig组合物;并通过使用本文所定义的基质的亲和层析法来清除组合物中可能含有的抗-A和/或抗-B抗体。
该基质的优点在于,其对抗-A及抗-B抗体具有高选择性及特异性,及其对抗-A及抗-B抗体高的结合容量。因此,该基质通过使得每个柱体积通过更大量的产品而可以在工业水平降低产品处理时间。
附图说明
图1为聚合物颗粒的简化示意图;根据本发明的特定实施方案,颗粒上移植了一种与A血型或B血型表位对应的寡糖。
图2为本发明优选实施方案的简化示意图;其描述了其上移植了与A血型表位对应的三糖的聚合物颗粒。
图3为本发明优选实施方案的简化示意图;说明了其上移植了与B血型表位对应的三糖的聚合物颗粒。
图4A为本发明特定实施方案的基质的简化示意图;该基质包括携带与A血型表位相对应的寡糖的颗粒,以及携带与B血型表位相对应的寡糖的颗粒的混合物。
图4B为本发明特定实施方案的基质的简化示意图;该基质包括既携带与A血型表位相对应的寡糖,又携带与B血型表位相对应的寡糖的颗粒。
图4C为本发明特定实施方案的基质的简化示意图;该基质包括携带与A血型表位相对应的寡糖的颗粒,携带与B血型表位相对应的寡糖的颗粒,以及既携带与A血型表位相对应的寡糖又携带与B血型表位相对应的寡糖的颗粒的混合物。
发明详述
介质:
本发明的基质包括聚合物颗粒的介质,并且优选呈凝胶形式或树脂形式。
这些聚合物颗粒优选呈球形或椭圆形,尤其可呈珠状。总体而言,这些颗粒的平均尺寸为约0.1μm至约1000μm,优选尺寸为约20至约500μm,还优选为约50至约200μm,还优选为约70μm至约120μm的直径。
优选地,这些颗粒呈多孔状。
聚合物可为天然或非天然的、有机或无机的、交联或非交联的聚合物。
聚合物优选为有机聚合物,优选为交联的聚合物。
在优选实施方案中,该聚合物为纤维素;该颗粒优选为多孔纤维素珠。
更优选地,该聚合物为交联的纤维素。
其他可能类型的聚合物包括琼脂糖、葡聚糖、聚丙烯酸酯、聚苯乙烯、聚丙烯酰胺、聚甲基丙烯酰胺、苯乙烯及二乙烯基苯的共聚物,或这些聚合物的混合物。
该颗粒可提供一种层析媒介,例如其可用于填充柱。
配体:
本发明介质所荷载的配体为代表A血型抗原或B血型抗原的寡糖,这些均为天然糖。
更特别地,本发明的基质所荷载的与A血型表位相对应的寡糖和/或与B血型表位相对应的寡糖通常为三糖。术语“与血型表位相对应的寡糖”指从抗原方面来看,具有分别被抗-A抗体及抗-B抗体识别的抗原决定簇的相同或类似的单位。本发明所述介质所荷载的配体因此则特异于抗-A抗体或抗-B抗体。
优选地,在本发明中当作配体使用的与A血型表位相对应的寡糖为三糖:N-乙酰半乳糖胺(GalNAc)-半乳糖(Gal)-岩藻糖,更特别是N-乙酰Galα1-3(Fucα1-2)Gal;结合间隔基,优选通过氧原子结合半乳糖位置1处的碳:
优选地,在本发明中当作配体使用的与B血型表位相对应的寡糖为寡糖:半乳糖-半乳糖-岩藻糖,更特别是Galα1–3(Fucα1–2)Gal;结合间隔基优选通过氧原子结合半乳糖位置1处的碳:
间隔基:
配体以共价方式与间隔基相连;间隔基连接配体及颗粒。间隔基能够减小空间位阻并提高配体与抗-A及抗-B抗体的可及性以结合。
间隔基的作用在于将与A血型表位相对应的寡糖(如前文所述,优选是N–乙酰Galα1–3(Fucα1–2)Gal的三糖)固定于颗粒上;或将与B血型表位相对应的寡糖(如前文所述,优选是Galα1–3(Fucα1–2)Gal的三糖)固定于颗粒上。
颗粒可具有若干间隔基。
举例而言,位于配体与间隔基之间的键可为酰胺键。
典型的间隔基至少包括一个C原子、O原子、N原子或S原子。
比如其可为-(CH2)mX(CH2)n-或-(CH2)mX1(CH2)nX2(CH2)p-,其中各X、X1及X2彼此独立地选自O、S、NH及共价键;而各m、n及p则分别单独选自0、1、2、3、4、5或6。在另一实施方案中,则可用OH基团和/或甲基的同等数量替代上述氢原子的1、2或3个。
在一个实施方案中,间隔基包含选自如下的结构:
其中各X1及X2均可单独选自O、S及NH;而各Ra、Rb、Rc及Rd均可单独选自H、OH及甲基。
在另一实施方案中,间隔基包含选自如下的结构:
在优选实施方案中,该间隔基具有式(I)-NH-R1-CO-NH-R2-;其中,R1为C4-C6烷基,R2为C3-C8烷基;并且该间隔基通过其胺官能团与颗粒结合(上文以粗体显示)。该基质的示意图已在图1中给出。
R1为直链或支链C4-C6烷基,优选直链C4-C6烷基。优选地,R1为C5烷基。
R2为直链或支链C3-C8烷基,优选直链C3-C8烷基。优选地,R2为C3烷基。
在优选实施方案中,配体(优选诸如前文所述的三糖)将通过于下式中所述的间隔基移植到颗粒上:(颗粒)-NH-C5H10-CO-NH-C3H6-(配体)。
在优选实施方案中,混合交联的纤维素珠;如图2中所述,将与A血型表位相对应的三糖(N-乙酰半乳糖胺(GalNAc)-半乳糖(Gal)-岩藻糖)的配体移植至该纤维素珠上;并如图3中所述,将与B血型表位相对应的三糖(半乳糖-半乳糖-岩藻糖)的配体移植至该纤维素珠上。可通过下式中的间隔基将三糖移植至珠上:(珠)-NH-C5H10-CO-NH-C3H6-(配体)。
制备基质:
配体由位于颗粒与间隔基之间以及间隔基与配体之间的共价键以化学方式进行固定。
可由本领域技术人员以常规方式进行固定。
在优选实施方案中,颗粒珠携带臂-NH-R1-COOH。优选地,其是ε–氨基己酸(其中R1为戊基)。
一般而言,该颗粒可使用双官能试剂加以激活,例如环氧氯丙烷、环氧溴丙烷、二溴丙醇及二氯丙醇、二溴丁烷、乙二醇二缩水甘油醚、丁二醇二缩水甘油醚、二乙烯基砜、烯丙基缩水甘油醚及溴化烯丙基。该双官能试剂既可与颗粒起反应,又可与臂-NH-R1-COOH起反应。诸如烯丙基溴的烯丙基杂官能团化合物优选为双官能试剂,且能够得到活性基质。
对于某些固体介质而言,例如纤维素,含有水凝胶的复合材料,或含羟基的其他材料,可在与双官能试剂发生反应之前,采用诸如氢氧化物的源头材料移除羟基内的质子。
随后可通过连接基团-NH-R2-将代表A血型抗原和/或B血型抗原的配体固定于携带臂-NH-R1-COOH的活化的颗粒上;其中R2为直链或支链C3-C8烷基,优选直链C3-C8烷基。就此,颗粒荷载的臂-NH-R1-COOH的COOH官能团将在使用N-乙氧羰基-2-乙氧基-1,2-二氢喹啉(EEDQ)型缩合剂的情况下与配体NH2-R2-寡糖中的NH2官能团发生反应。
本领域技术人员可将代表A血型抗原的配体,或代表B血型抗原的配体移植至同一颗粒上,或将二者共同移植至同一颗粒上。优选地,所用的间隔基为相同间隔基。如此便可有利于保证将代表A血型抗原的配体以及代表B血型抗原的配体进行同等移植。本领域技术人员此外根据配体及颗粒的反应性情况知晓如何调整合适的条件,以便获得携带确定配体比例的颗粒,所述配体代表A血型抗原以及代表B血型抗原。
呈凝胶状的基质可以通过标准的在携带配体的聚合物颗粒中添加缓冲剂制备而成;此法已为本领域技术人员熟知,以使得可获得适用于亲和层析方法的凝胶状基质。
有利地,配体密度(即每个基质体积内移植的配体的量,所述配体即与A血型表位或B血型表位相对应的寡糖)包含在约0.2至约0.7mg/ml的基质之间,优选在约0.3至约0.4mg/ml基质之间。优选地,寡糖的密度为约0.3mg/ml的基质。
根据本发明,与1mg/ml的密度(例如在申请WO2007/077365中所述的,来自GlycorexTransplantationAB(瑞典)的凝胶GLYCOSORB上的密度)相比,该密度能够显著降低以亲和层析法进行纯化所需的时间。可增加基质体积;而且,亦可增加通过层析柱的流量。因此,在配体密度为0.3mg/ml的基质时(与1mg/ml的密度相比),可将方法所需时间减少一半,这在工业化生产中表示显著的增益。有利地,降低配体的密度同样能够降低基质的成本,从而使得在根据本发明的基质上的亲和步骤成本较低,从而有助于降低最终免疫球蛋白组成物的价格成本。有利地,降低配体密度的同时,能够保持清除抗-A和/或抗-B抗体的能力。
在优选实施方案中,那么可以混合其中移植了至少一种与A血型表位相对应的寡糖的聚合物颗粒,和其中移植了至少一种与B血型表位相对应的寡糖的聚合物颗粒,例如按25/75至75/25(v/v)的比例进行混合,优选按约50/50(v/v)的比例进行混合。
必要时,也可以将其中移植了至少一种与A血型表位相对应的寡糖的聚合物颗粒,其中移植了至少一种与B血型表位相对应的寡糖的聚合物颗粒与这样的聚合物颗粒混合,该颗粒即携带了与A血型表位相对应的寡糖,也携带了与B血型表位相对应的寡糖。
亲和层析:
此处所定义的基质可在结合抗-A和/或抗-B抗体的亲和层析过程中使用。
就此,可将基质加入至层析柱内。在进行工业化制备的过程中,柱容量可为1至150L,甚至250至500L,视需要而定。在指导规模应用的情况下,可使用高度为1至50cm的层析柱;其直径则与所用柱的高度相匹配。可根据与溶液中抗-A和/或抗-B抗体的原始比例相比意欲达到的抗-A和/或抗-B抗体的残余比例来对柱中所用基质的体积进行调整。同样可对基质的体积进行调整,使其符合工业方法的相关要求,尤其需要符合待处理产品体积方面的要求。
需要将免疫球蛋白液体制品(或血液产品衍生物)中所存在的抗-A和/或抗-B抗体结合至基质中;并回收排除了抗-A和/或抗-B抗体的未吸收的产品。在可使抗体与基质所荷载的配体结合的速率和条件下,将免疫球蛋白液体制品或血液产品衍生物灌注通过基质。
基质可反复再利用,如可再利用约100次而不受损。可由本领域技术人员采用已知方法对其进行再生,例如通过使用如1M的氢氧化钠(NaOH)的处理再生。再生后的再利用符合医药制造工业方法的安全要求,尤其符合移除生物污染以及消除将导致批次之间污染的痕量产品方面的相关要求。对亲和基质进行再利用的优点在于,能够在制造免疫球蛋白的过程中降低工业成本。
因此,根据本发明所述的亲和基质可良好适用于制备诸如免疫球蛋白的药物的工业应用,且可完全且充分地移除溶液中最初所含的抗-A和/或抗-B抗体,而不明显地增加最终免疫球蛋白产品的工业成本。
免疫球蛋白产品纯化:
使用本发明所述基质的亲和层析法特别有利于纯化免疫球蛋白制品或组合物,以清除在这些制品或组合物中可能存在的不需要的抗-A和/或抗-B抗体,并因此制备具有治疗用途的免疫球蛋白浓缩物。
“清除”可以理解为实质性地降低所出现的抗-A和/或抗-B抗体的量,优选降低至少25%,更优选降低至少为50%,还更优选降低至少为80%或90%。根据本发明在亲和层析方法之后所获得之Ig浓缩物,其各自抗-A及抗-B抗体含量符合Coombs直接测试的阴性检测方面(稀释至1/64)的要求;而其IgG原始浓度为30g/l。优选地,在亲和层析方法之后,通过本发明所获得的Ig浓缩物包含不大于23ng/mgIgG的抗-A抗体,和不大于20ng/mgIgG的抗-B抗体。定量剩余抗-A及抗-B抗体的方法已尤其在申请WO2007/077365中予以说明。优选地,可采用流式细胞术测定抗-A及抗-B抗体剂量;其原理为根据意欲达到的抗-A及抗-B抗体方面的特殊滴度测定值来使用A型或B型人体红细胞,并使用与这些抗体的含量成比例的萤光信号测量值。总体而言,以流式细胞术测定剂量的方法需要使用诸如免疫球蛋白阳性对照物(例如1:32滴度的EDQM,参考号07/306;Thorpe等人2009,VoxSang.97,160-168)的标准样品或单克隆抗-D抗体浓缩物或其他各种相应的参考产品。
免疫球蛋白产品主要含有IgG。这些IgG通常是多克隆IgG,其从血浆获得或者从已经富含Ig的血浆级分中获得。
用于治疗用途的Ig浓缩物,其浓度为约50至100g/l。这些浓缩物专门用于临床用途,尤其用于进行静脉注射。就此,该产品应保障其安全性;在必要时需要含有诸如可与临床用途相匹配的赋形剂,例如稳定剂。
用于治疗用途的Ig浓缩物,同样可以皮下给药。在此情况下,产品浓度将大于或等于100g/l,有利地大于或等于150g/l。
用于治疗用途的Ig浓缩物,同样可以肌内途径给药。
可以如下方式获得Ig浓缩物:
a)采用诸如乙醇分级和/或辛酸分级的方法和/或层析分离法来制备Ig组合物;
b)以在本发明所述基质上对Ig组合物进行渗滤的方式来进行免疫亲和层析;
以及c)生物安全化步骤;优选使用纳米过滤来移除污染性病毒和/或颗粒。
可以采用Cohn等人首创之乙醇分级法(Cohn等人,1946;J.Am.Chem.Soc.68,459;Oncley等人,1949;J.Am.Chem.Soc.71,541)或诸如申请EP0703922及WO99/64462中所述的层析分离法来获得Ig组合物,或以Steinbuch等人1969,ArchBiochemBiophys.134(2):279-84中所述的辛酸分级法来获得Ig组合物。最优选使用由申请人在专利申请WO94/29334及WO02/092632中开发的方法,以及特别最优选在申请WO02/092632中特别说明的方法。在此情况下,将血浆或富含IgG的血浆级分进行辛酸分级法(通过沉淀非免疫球蛋白污染物进行预纯化),单独在PH值呈碱性的阴离子交换剂树脂介质上进行层析,采用合适的缓冲液在PH值介于4与7之间的步骤中选择性洗脱IgG。可以进行病毒灭活处理,例如通过采用Horowitz在专利US4764369中所述的溶剂-去污剂来进行。
由此所获得的IgG级分已经得到浓缩,但随后需要通过超滤及无菌过滤的方式进行额外的浓缩步骤。
随后将该浓缩物在本发明基质上实施免疫性亲和层析步骤。优选地,在进行层析前,将免疫球蛋白制品的PH值调整至约6至约7的范围,优选pH值为约6。
层析柱的荷载与抗-A和/或抗-B抗体的理想剩余率相匹配。此类基质之特异性在于,无需提前确定IgG级分的条件,即通过血浆分级技术获得的各类IgG级分或浓缩物都是适合的。
无需使用洗脱设备便可对浓缩物进行渗滤。因此,无论以何种方式获得IgG浓缩物,均可渗滤通过层析柱,必要时可使用泵来进行渗滤。该渗滤方式将滞留抗-A及抗-B抗体。配体与Ig制品之间的接触时间将大于或等于一分钟,有利地为2分钟的级别。
随后可以用水来清洗层析柱,以收集仍存在于层析柱无效腔内的IgG。
在对IgG浓缩物进行渗滤后,可以获得贫化了抗-A及抗-B抗体的IgG级分。
该方法随后还包括以超滤及无菌过滤的方式进行的浓缩步骤。
随后可以对层析柱和基质进行洗涤和洗脱,以便于解吸所滞留的抗-A及抗-B抗体。
本发明中随后所举之实施例并不起限定其保护范围的作用。
实施例:亲和层析(工业规模)
层析条件:
根据申请WO2002/092632中所描述的方法以分级方式获得的免疫球蛋白制品,包含10±2g/l的IgG,对其在层析柱上进行工业规模的抗-A/抗-B亲和层析步骤,所述层析柱包括交联的纤维素珠的50/50(v/v)混合物;其中,一种珠上移植了与A血型表位相对应的三糖(N-乙酰半乳糖胺(GalNAc)-半乳糖(Gal)-岩藻糖),如图2所示,凝胶用“IsoAHyperCel”指代;另一种交联的纤维素珠上移植了与B血型表位相对应的三糖(半乳糖-半乳糖-岩藻糖),如图3所示,凝胶用“IsoBHyperCel”指代。
将采用下式中所述的间隔基将三糖移植至珠上:
(珠)-NH-C5H10-CO-NH-C3H6-(配体)
移植的三糖的密度为0.3mg/ml的基质凝胶。
为核实抗-A及抗-B残余活性,以洗出液来指代免疫球蛋白制品(经过阴离子交换剂层析方法后获得的级分),将其pH值调节至6(±0.05);该制品通过1ml的层析柱(包括0.5ml的IsoAHyperCel凝胶+0.5ml的IsoBHyperCel凝胶)。
层析柱规格:D=0.5cm×5.1cm(层析柱体积(VC)=1ml)。
接触时间:2分钟
荷载:6g洗出液制品/mg的A+B配体,即每根层析柱中180ml产品。
试验后,用水(至少10VC)洗涤凝胶。
方法的各个步骤的条件已在下表中予以概述:
步骤表:
VC:层析柱体积;NAF:未被吸收的级分
收集单一的未被吸收的级分(NAF)。
剂量测试:
通过定量洗涤及渗滤后未被吸收的级分中的IgG剂量来测定其产量。根据下述技术,通过流式细胞术(与欧洲药典中所要求的稀释技术相比,此技术更敏感及精确)来测定抗-A及抗-B残余活性,其对应于最终制品中所存在的抗-A及抗-B同族凝集素浓度与其在进行亲和层析步骤前的浓度的百分比。
A血型及B血型的红细胞可在EDTA上进行收集,并采用浓度为0.9%的NaCl溶液洗涤两次(在2次洗涤之间以1730g离心5分钟)。2.106个红细胞分布于微滴定盘上,添加50μL的标准阳性对照物(1:32EDQM,参考号07/306;Thorpe等人2009)或添加稀释至工作浓度的样品(PBSpH7.4,1%BSA)。将微滴定盘在37℃的搅拌条件下温育2小时。洗涤后,使用标记有藻红蛋白(PE)的在PBS-BSA内稀释至1/20的人抗IgG山羊F(ab')2抗体(Fc特异的)(BeckmanCoulter)。在室温下以避光方式将微滴定盘温育30分钟。
洗涤后,将各沉淀物再悬浮于200μLPBS-BSA中,并在流式细胞仪(BeckmanCoulterCytomicsFC500)上读取其读数。
相对于IgG的浓度(其浓度为0.23g/L至30g/L)报告阳性对照的平均萤光强度(MFI)(标准曲线)。其结果采用样品的斜率与阳性标准的斜率之间的比值来表示。标准曲线方程式为y=ax+b;其中“a”为标准曲线的斜率值;而“b”为与试验基础噪音相对应的零点。由于样本的方程式为y'=a'x+b,并且使用已知的样本的MFI值(y')及IgG的浓度(x'),因此可采用式[(MFI-b)/[IgG浓度]]/a来计算其斜率比值。
结果:
所获得的IgG产量以及抗-A及抗-B残余活性已在下表中列出:
结果表:
该表的结果表明,在未被吸收的级分中IgG的产量为98%,此证明亲和介质在抗-A及抗-B同族凝集素方面具有良好特异性。
该表的结果同样表明,抗-A及抗-B残余活性实质性降低,分别为12%及8%。因此获得的产品符合在IgG初始浓度为30g/L的情况下所进行的Coombs直接测试(稀释至1/64)的阴性结果方面的要求。
Claims (17)
1.一种凝胶形式的亲和层析基质,其包括聚合物颗粒,在这些颗粒上移植了至少一种与A血型和/或B血型表位相对应的寡糖;该寡糖通过间隔基移植至该颗粒上,其特征在于:寡糖的密度包含在0.2至0.7mg/ml的基质之间,且所述间隔基具有式(I)-NH-R1-CO-NH-R2-,其中,R1为C4-C6烷基,R2为C3-C8烷基,且所述间隔基通过其胺官能团与颗粒结合。
2.根据权利要求1的基质,包含(i)聚合物颗粒,其上移植了至少一种与A血型表位相对应的寡糖;和/或(ii)聚合物颗粒;其上移植了至少一种与B血型表位相对应的寡糖。
3.根据权利要求2的基质,其包括(i)聚合物颗粒,其上移植了至少一种与A血型表位相对应的寡糖;以及(ii)聚合物颗粒;其上移植了至少一种与B血型表位相对应的寡糖。
4.根据权利要求1的基质,其包括聚合物颗粒,其上即移植了至少一种与A血型表位相对应的寡糖,还移植了至少一种与B血型表位相对应的寡糖。
5.根据权利要求1的基质,其包括如下物质的混合物:(i)聚合物颗粒,其上移植了至少一种与A血型表位相对应的寡糖;(ii)聚合物颗粒,其上移植了至少一种与B血型表位相对应的寡糖;以及(iii)聚合物颗粒,其上即移植了至少一种与A血型表位相对应的寡糖,还移植了至少一种与B血型表位相对应的寡糖。
6.根据权利要求1至5中任一项的基质,其中寡糖的密度包含在0.3至0.4mg/ml的基质之间,优选密度为约0.3mg/ml的基质。
7.根据权利要求1至6中任一项的基质,其中与A血型表位相对应的寡糖和/或与B血型表位相对应的寡糖为三糖。
8.根据权利要求7的基质,其中与A血型表位相对应的寡糖为N-乙酰半乳糖胺(GalNAc)-半乳糖(Gal)-岩藻糖的三糖。
9.根据权利要求7的基质,其中与B血型表位相对应的寡糖为半乳糖-半乳糖-岩藻糖的寡糖。
10.根据权利要求1至9中任一项的基质,其中R1为C5烷基。
11.根据权利要求1至10中任一项的基质,其中R2为C3烷基。
12.根据权利要求3的基质,其中移植了至少一种与A血型表位相对应的寡糖的聚合物颗粒与移植了至少一种与B血型表位相对应的寡糖的聚合物颗粒按25/75至75/25(v/v)的比例进行混合,优选按约50/50(v/v)的比例进行混合。
13.根据权利要求1至12中任一项的基质,其中聚合物为交联的聚合物。
14.根据权利要求13的基质,其中该聚合物为纤维素,并且该颗粒优选为多孔纤维素珠。
15.权利要求1至14中任一项的基质在结合抗-A抗体和/或抗-B抗体的亲和层析方法中的应用。
16.根据权利要求15的基质的应用,其用于在工业规模制备用于治疗用途的免疫球蛋白G(IgG)。
17.一种制备用于治疗用途的免疫球蛋白G(IgG)浓缩物的方法,包括通过乙醇分级和/或辛酸分级的方法和/或层析分离法来从血浆中获得Ig组合物;并通过采用根据权利要求1-14中任一项的基质的亲和层析法来清除可能存在于组合物中的抗-A和/或抗-B抗体。
Applications Claiming Priority (5)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
FR1356635 | 2013-07-05 | ||
FR1356636 | 2013-07-05 | ||
FR1356636A FR3008098B1 (fr) | 2013-07-05 | 2013-07-05 | Matrice de chromatographie d'affinite a densite de ligands reduite |
FR1356635A FR3008097B1 (fr) | 2013-07-05 | 2013-07-05 | Matrice de chromatographie d'affinite |
PCT/FR2014/051734 WO2015001277A1 (fr) | 2013-07-05 | 2014-07-04 | Matrice de chromatographie d'affinité |
Publications (1)
Publication Number | Publication Date |
---|---|
CN105358228A true CN105358228A (zh) | 2016-02-24 |
Family
ID=51303088
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201480038362.8A Pending CN105358228A (zh) | 2013-07-05 | 2014-07-04 | 亲和层析基质 |
Country Status (13)
Country | Link |
---|---|
US (1) | US10611826B2 (zh) |
EP (1) | EP3016729B1 (zh) |
JP (1) | JP2016532100A (zh) |
KR (1) | KR20160029840A (zh) |
CN (1) | CN105358228A (zh) |
AU (1) | AU2014285971A1 (zh) |
CA (1) | CA2916566A1 (zh) |
ES (1) | ES2793176T3 (zh) |
IL (1) | IL243414A0 (zh) |
MX (1) | MX2016000063A (zh) |
PL (1) | PL3016729T3 (zh) |
TW (1) | TW201509432A (zh) |
WO (1) | WO2015001277A1 (zh) |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN108101981A (zh) * | 2018-01-15 | 2018-06-01 | 四川远大蜀阳药业股份有限公司 | 一种静注免疫球蛋白的生产工艺 |
CN113166233A (zh) * | 2018-12-05 | 2021-07-23 | 西托索尔本茨公司 | 用于从人血浆和全血中去除抗a和/或抗b抗体的基于交联多糖的吸收剂 |
Families Citing this family (12)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP6189751B2 (ja) | 2010-12-30 | 2017-08-30 | ラボラトワール フランセ デュ フラクショヌマン エ デ ビオテクノロジーLaboratoire Francais du Fractionnement et des Biotechnologies | 病原体不活性化剤としてのグリコール |
CN105308068A (zh) | 2013-02-13 | 2016-02-03 | 法国化学与生物科技实验室 | 高度半乳糖基化的抗TNF-α抗体及其用途 |
BR112015019348A2 (pt) | 2013-02-13 | 2017-08-22 | Lab Francais Du Fractionnement | Métodos para produção de proteína com glicosilação modificada e com sialilação aumentada, para aumentar a atividade de sialil transferase na glândula mamária e para produzir sialil transferase, proteína com glicosilação modificada ou proteína com sialilação aumentada, composição, sialil transferase, mamífero transgênico, e, célula epitelial mamária |
PL3016729T3 (pl) | 2013-07-05 | 2020-09-07 | Laboratoire Français Du Fractionnement Et Des Biotechnologies Société Anonyme | Matryca do chromatografii powinowactwa |
FR3026950A1 (fr) * | 2014-10-09 | 2016-04-15 | Lab Francais Du Fractionnement | Procede de preparation de plasma universel |
US20170066839A1 (en) | 2015-09-08 | 2017-03-09 | Merck Patent Gmbh | Novel affinity chromatography media for removal of anti-a and/or anti-b antibodies |
US10697983B2 (en) | 2015-09-08 | 2020-06-30 | Merck Patent Gmbh | Methods of evaluating quality of media suitable for removing anti-A or anti-B antibodies |
US10697982B2 (en) | 2015-09-08 | 2020-06-30 | Merck Patent Gmbh | Methods of evaluating quality of a chromatography media which binds anti-A or anti-B antibodies |
US11660550B2 (en) | 2017-03-07 | 2023-05-30 | Jmva Biotech Ab | Modified adsorptive surfaces |
JP7026116B2 (ja) * | 2017-08-23 | 2022-02-25 | Jsr株式会社 | クロマトグラフィー用担体、リガンド固定担体、クロマトグラフィーカラム、標的物質の精製方法、及びクロマトグラフィー用担体の製造方法 |
WO2024091527A1 (en) * | 2022-10-25 | 2024-05-02 | Donaldson Company, Inc. | Separation media and purification methods for carbohydrate containing molecules using the same |
WO2024091525A1 (en) * | 2022-10-25 | 2024-05-02 | Donaldson Company, Inc. | Separation media and purification methods for blood antibodies using the same |
Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1196688A (zh) * | 1996-06-20 | 1998-10-21 | 巴克斯特国际有限公司 | 亲和性膜系统及其应用方法 |
US6069236A (en) * | 1993-06-14 | 2000-05-30 | Association Pour L'essor De La Transfusion Sanguine Dans La Region Du Nord | Immunoglobulin G concentrate for therapeutic use and process for producing said concentrate |
US20040132979A1 (en) * | 2001-05-11 | 2004-07-08 | Chtourou Abdessatar Sami | Method for preparing human immunoglobulin concentrates for therapeutic use |
CN101098722A (zh) * | 2005-02-17 | 2008-01-02 | 弗雷森纽斯医疗护理德国有限责任公司 | 从液体尤其是血液中除去物质的装置 |
CN101346153A (zh) * | 2005-12-26 | 2009-01-14 | 分馏和生物工艺法国实验室公司 | 除去抗A、抗B抗体以及多反应性免疫球蛋白IgG的免疫球蛋白G(IgG)浓缩物 |
WO2013066251A1 (en) * | 2011-10-30 | 2013-05-10 | Glycorex Ab | Method for the reduction or elimination of one or more components from a blood product |
Family Cites Families (119)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US3519558A (en) | 1968-02-15 | 1970-07-07 | Abcor Inc | Separation of similar solutes using a semipermeable membrane |
BE759625A (fr) | 1969-12-10 | 1971-04-30 | Molkereigenossenschaft Dahlenb | Procede et dispositif pour la dialyse du lait, en particulier pour l'obtention de l'albumine du lait et du petit-lait |
CH566798A5 (zh) | 1973-05-29 | 1975-09-30 | Battelle Memorial Institute | |
SE396017B (sv) | 1974-12-23 | 1977-09-05 | Alfa Laval Ab | Filtreringsforfarande, serskilt for ultrafiltrering |
US4351710A (en) | 1980-01-10 | 1982-09-28 | Ionics, Incorporated | Fractionation of protein mixtures |
US4350156A (en) | 1980-05-29 | 1982-09-21 | Japan Foundation For Artificial Organs | Method and apparatus for on-line filtration removal of macromolecules from a physiological fluid |
JPS583705B2 (ja) | 1980-07-18 | 1983-01-22 | 川澄化学工業株式会社 | 二重ロ過型血漿分離交換装置 |
US4420398A (en) | 1981-08-13 | 1983-12-13 | American National Red Cross | Filteration method for cell produced antiviral substances |
US4764369A (en) | 1983-07-14 | 1988-08-16 | New York Blood Center Inc. | Undenatured virus-free biologically active protein derivatives |
US4888115A (en) | 1983-12-29 | 1989-12-19 | Cuno, Incorporated | Cross-flow filtration |
CA1244349A (en) | 1985-06-26 | 1988-11-08 | Jen-Chang Hsia | Purification of hemoglobin and modified hemoglobin by affinity chromatography |
US4789482A (en) | 1986-02-10 | 1988-12-06 | Millipore Corporation | Method for separating liquid compositions on the basis of molecular weight |
DE3751873T2 (de) | 1986-04-09 | 1997-02-13 | Genzyme Corp | Genetisch transformierte Tiere, die ein gewünschtes Protein in Milch absondern |
US6727405B1 (en) | 1986-04-09 | 2004-04-27 | Genzyme Corporation | Transgenic animals secreting desired proteins into milk |
DE3876273T2 (de) | 1987-04-11 | 1993-05-27 | Ciba Geigy Ag | Isoelektrisches fokussierverfahren sowie einrichtung zur durchfuehrung dieses verfahrens. |
SE464687B (sv) * | 1987-11-10 | 1991-06-03 | Biocarb Ab | Foerfarande foer framstaellning av en gelprodukt |
US5948441A (en) | 1988-03-07 | 1999-09-07 | The Liposome Company, Inc. | Method for size separation of particles |
US5256294A (en) | 1990-09-17 | 1993-10-26 | Genentech, Inc. | Tangential flow filtration process and apparatus |
US5648253A (en) | 1990-12-20 | 1997-07-15 | Tsi Corporation | Inhibitor-resistant urokinase |
US6448469B1 (en) | 1991-10-02 | 2002-09-10 | Genzyme Corporation | Production of membrane proteins in the milk of transgenic nonhuman mammals |
JP3801196B2 (ja) | 1993-03-09 | 2006-07-26 | ジェンザイム・コーポレイション | 乳からの対象化合物の単離 |
DE4326665C2 (de) | 1993-08-09 | 1995-07-13 | Biotest Pharma Gmbh | Verfahren zur Sterilfiltration von Milch |
US5827690A (en) | 1993-12-20 | 1998-10-27 | Genzyme Transgenics Corporatiion | Transgenic production of antibodies in milk |
US5518624A (en) | 1994-05-06 | 1996-05-21 | Illinois Water Treatment, Inc. | Ultra pure water filtration |
US5843705A (en) | 1995-02-21 | 1998-12-01 | Genzyme Transgenic Corporation | Transgenically produced antithrombin III |
US5597486A (en) | 1995-05-01 | 1997-01-28 | Millipore Investment Holdings Limited | Membrane filtration with optimized addition of second liquid to maximize flux |
EP0871671A1 (en) | 1995-09-07 | 1998-10-21 | PPL Therapeutics (Scotland) Limited | Purification of alpha-1 proteinase inhibitor |
US6054051A (en) | 1996-01-17 | 2000-04-25 | Genentech, Inc. | Tangential-flow filtration system |
US6268487B1 (en) | 1996-05-13 | 2001-07-31 | Genzyme Transgenics Corporation | Purification of biologically active peptides from milk |
US6686457B1 (en) * | 1996-12-23 | 2004-02-03 | Kurt Nilsson | Material |
CA2281949A1 (en) | 1997-02-25 | 1998-08-27 | Genzyme Transgenics Corporation | Transgenically produced non-secreted proteins |
US6210736B1 (en) | 1997-06-17 | 2001-04-03 | Genzyme Transgenics Corporation | Transgenically produced prolactin |
BR9812945A (pt) | 1997-10-20 | 2000-08-08 | Genzyme Transgenics Corp | Sequências de ácido nucléico modificadas e processos para aumentar os nìveis de mrna e expressão de sistemas celulares |
RU2197500C2 (ru) | 1998-06-09 | 2003-01-27 | Статенс Серум Институт | Способ получения иммуноглобулинов для внутривенного введения и другие иммуноглобулиновые продукты |
US6548653B1 (en) | 1998-06-15 | 2003-04-15 | Genzyme Transgenics Corporation | Erythropoietin analog-human serum albumin fusion |
US20050181482A1 (en) | 2004-02-12 | 2005-08-18 | Meade Harry M. | Method for the production of an erythropoietin analog-human IgG fusion proteins in transgenic mammal milk |
US20030005468A1 (en) | 1998-06-19 | 2003-01-02 | Meade Harry M. | Methods and vectors for improving nucleic acid expression |
US20040117863A1 (en) | 1998-09-18 | 2004-06-17 | Edge Michael D. | Transgenically produced fusion proteins |
EP1818397A1 (en) | 1998-11-02 | 2007-08-15 | Trustees Of Tufts College | Methods for cloning animals |
US20030177513A1 (en) | 1998-11-02 | 2003-09-18 | Yann Echelard | Transgenic and cloned mammals |
US6580017B1 (en) | 1998-11-02 | 2003-06-17 | Genzyme Transgenics Corporation | Methods of reconstructed goat embryo transfer |
US7208576B2 (en) | 1999-01-06 | 2007-04-24 | Merrimack Pharmaceuticals, Inc. | Non-glycosylated human alpha-fetoprotein, methods of production, and uses thereof |
CA2361545C (en) | 1999-02-22 | 2009-01-27 | Henry Kopf | Purification of biological substances |
CA2362565A1 (en) | 1999-02-22 | 2000-08-31 | Eric F. Bernstein | Compositions and methods for prevention of photoaging |
WO2001026455A1 (en) | 1999-10-14 | 2001-04-19 | Genzyme Transgenics Corporation | Methods of producing a target molecule in a transgenic animal and purification of the target molecule |
US7544500B2 (en) | 1999-11-13 | 2009-06-09 | Talecris Biotherapeutics, Inc. | Process for the production of a reversibly inactive acidified plasmin composition |
AU5946501A (en) | 2000-05-05 | 2001-11-20 | Genzyme Transgenics Corp | Transgenically produced decorin |
NZ523220A (en) | 2000-06-19 | 2005-03-24 | Gtc Biotherapeutics Inc | Transgenically produced platelet derived growth factor |
EP1315418A4 (en) | 2000-08-10 | 2004-01-14 | Gtc Biotherapeutics Inc | SPERM CRYOPRESERVATION |
CA2424977C (en) | 2000-10-06 | 2008-03-18 | Kyowa Hakko Kogyo Co., Ltd. | Process for purifying antibody |
US20040226052A1 (en) | 2000-10-13 | 2004-11-11 | Meade Harry M. | Methods of producing a target molecule in a transgenic animal and purification of the target molecule |
US20030036637A1 (en) | 2001-06-13 | 2003-02-20 | Scott Fulton | Purification of human serum albumin |
US7651686B2 (en) | 2001-10-09 | 2010-01-26 | Mayo Foundation For Medical Education And Research | Enhancement of immune responses by 4-1bb-binding agents |
PL368755A1 (en) | 2001-11-28 | 2005-04-04 | Sandoz Ag | Chromatographic purification of recombinant human erythropoietin |
US20040133931A1 (en) | 2003-01-08 | 2004-07-08 | Gavin William G. | Method and system for fusion and activation following nuclear transfer in reconstructed embryos |
EP1463802A1 (en) | 2002-01-11 | 2004-10-06 | GTC Biotherapeutics, Inc. | Method and system for fusion and activation following nuclear transfer in reconstructed embryos |
ES2319636T3 (es) | 2002-02-05 | 2009-05-11 | Genentech, Inc. | Purificacion de proteinas. |
AU2003230725A1 (en) | 2002-04-01 | 2003-10-20 | Gtc Biotherapeutics, Inc. | A method for selecting cell lines to be used for nuclear transfer in mammalian species |
KR20040105838A (ko) | 2002-04-01 | 2004-12-16 | 지티씨바이오쎄라퓨틱스,인크. | 폐 질환의 치료 방법 |
PT1501369E (pt) | 2002-04-26 | 2015-09-21 | Genentech Inc | Purificação de proteínas com base na não afinidade |
CA2492561A1 (en) | 2002-07-15 | 2004-01-22 | Mayo Foundation For Medical Education And Research | Treatment and prophylaxis with 4-1bb-binding agents |
EP1534065A4 (en) | 2002-08-01 | 2005-11-09 | Gtc Biotherapeutics Inc | METHOD FOR RAPIDLY SELECTING LINES OF PRIMARY HOMOZYGOT CELLS TO PRODUCE TRANSGENIC ANIMALS BY NUCLEAR TRANSFER OF SOMATIC CELLS |
WO2004026427A2 (en) | 2002-09-17 | 2004-04-01 | Gtc Biotherapeutics, Inc. | Isolation of immunoglobulin molecules that lack inter-heavy chain disulfide bonds |
US7264728B2 (en) | 2002-10-01 | 2007-09-04 | Dow Corning Corporation | Method of separating components in a sample using silane-treated silica filter media |
US20040102380A1 (en) | 2002-11-18 | 2004-05-27 | Fulton Scott P. | Method for continuous, automated blending of solutions from acids and bases |
US7087719B2 (en) | 2002-11-19 | 2006-08-08 | Gtc Biotherapeutics, Inc. | Method for the crystallization of human serum albumin |
EP1565564A4 (en) | 2002-11-27 | 2006-06-07 | Gtc Biotherapeutics Inc | STILK-STABILIZED ANTIBODIES PRODUCED IN MILK AND METHOD FOR THE MANUFACTURE THEREOF |
US20040148648A1 (en) | 2002-12-10 | 2004-07-29 | Esmail Behboodi | Method and system for utilizing somatic cell nuclear transfer embryos as cell donors for additional nuclear transfer |
EP1601788A4 (en) | 2003-02-24 | 2006-11-15 | Gtc Biotherapeutics Inc | TANGENTIAL FILTRATION METHODS AND APPARATUS THEREFOR |
CN1871252A (zh) | 2003-09-05 | 2006-11-29 | Gtc生物治疗学公司 | 在转基因哺乳动物奶中生产融合蛋白的方法 |
CA2538722A1 (en) | 2003-09-15 | 2005-04-28 | Gtc Biotherapeutics, Inc. | Expression of dominant negative transmembrane receptors in the milk of transgenic animals |
CA2543193C (en) | 2003-10-27 | 2015-08-11 | Wyeth | Removal of high molecular weight aggregates using hydroxyapatite chromatography |
US20050169908A1 (en) | 2004-01-23 | 2005-08-04 | Kazunori Murakami | Use of aerosolized antithrombin to treat acute lung injury |
US20050186608A1 (en) | 2004-02-19 | 2005-08-25 | Olsen Byron V. | Method for the production of transgenic proteins useful in the treatment of obesity and diabetes |
US20050192226A1 (en) | 2004-02-20 | 2005-09-01 | Perenlei Enkhbaatar | Method of preventing fibrin clots in pulmonary tissue through the use of aerosolized anticoagulants |
US20050197496A1 (en) | 2004-03-04 | 2005-09-08 | Gtc Biotherapeutics, Inc. | Methods of protein fractionation using high performance tangential flow filtration |
US20050245444A1 (en) | 2004-04-30 | 2005-11-03 | Yann Echelard | Method of using recombinant human antithrombin for neurocognitive disorders |
US20060121004A1 (en) | 2004-12-07 | 2006-06-08 | Yann Echelard | Methods of reducing the incidence of rejection in tissue transplantation through the use of recombinant human antithrombin |
US20060123500A1 (en) | 2004-12-07 | 2006-06-08 | Gtc Biotherapeutics, Inc. | Methods of prescreening cells for nuclear transfer procedures |
US20060130159A1 (en) | 2004-12-09 | 2006-06-15 | Nick Masiello | Method of purifying recombinant MSP 1-42 derived from Plasmodium falciparum |
US20060168671A1 (en) | 2005-01-21 | 2006-07-27 | Gavin William G | Injection of caprine sperm factor (cSF), phospholipase C zeta (PLCzeta) and adenophostin A as alternative methods of activation during nuclear transfer in the caprine species |
US20080019905A9 (en) | 2005-02-18 | 2008-01-24 | Strome Scott E | Method of using an anti-CD137 antibody as an agent for radioimmunotherapy or radioimmunodetection |
US20060182744A1 (en) | 2005-02-15 | 2006-08-17 | Strome Scott E | Anti-CD137 antibody as an agent in the treatment of cancer and glycosylation variants thereof |
US20060286548A1 (en) | 2005-06-16 | 2006-12-21 | Gregory Liposky | Method of making recombinant human antibodies for use in biosensor technology |
MY144484A (en) | 2005-06-17 | 2011-09-30 | Wyeth Corp | Methods of purifying anti a beta antibodies |
US20070037192A1 (en) | 2005-07-25 | 2007-02-15 | Gtc Biotherapeutics, Inc. | Method of purifying recombinant human antithrombin to enhance the viral and prion safety profile |
EP2388272A1 (en) | 2005-10-21 | 2011-11-23 | GTC Biotherapeutics, Inc. | Antibodies with enhanced antibody-dependent cellular cytoxicity activity, methods of their production and use |
AU2013200440B2 (en) * | 2005-12-26 | 2016-03-31 | Laboratoire Francais Du Fractionnement Et Des Biotechnologies | Immunoglobulin G (IgG) concentrate depleted of anti-A and anti-B antibodies and of polyreactive IgGs |
US7531632B2 (en) | 2006-02-16 | 2009-05-12 | Gtc Biotherapeutics, Inc. | Clarification of transgenic milk using depth filtration |
US8173860B2 (en) | 2006-04-21 | 2012-05-08 | Gtc Biotherapeutics, Inc. | Non-human transgenic mammal expressing a human FcRn on its mammary gland cells and expressing a transgenic protein-human Fc-domain fusion |
FR2901796A1 (fr) | 2006-05-31 | 2007-12-07 | Lab Francais Du Fractionnement | Procede d'extraction d'une ou de plusieurs proteines presentes dans du lait |
WO2008073620A2 (en) | 2006-11-02 | 2008-06-19 | Neose Technologies, Inc. | Manufacturing process for the production of polypeptides expressed in insect cell-lines |
EP2144681A4 (en) * | 2007-05-04 | 2011-10-05 | Kurt G I Nilsson | MATERIAL FOR THE DEPOSITION OF A BIOMOLECULAR |
WO2009134389A2 (en) | 2008-05-01 | 2009-11-05 | Gtc Biotherapeutics, Inc. | An anti-cd137 antibody as an agent in the treatment of inflammatory conditions |
JP5155453B2 (ja) | 2008-09-15 | 2013-03-06 | イー・エム・デイー・ミリポア・コーポレイシヨン | タンパク質をベースとするアフィニティークロマトグラフィー樹脂からのタンパク質漏出を定量するための方法 |
US20110082083A1 (en) | 2009-04-10 | 2011-04-07 | Gtc Biotherapeutics, Inc. | Formulations of liquid stable antithrombin |
JP6189751B2 (ja) | 2010-12-30 | 2017-08-30 | ラボラトワール フランセ デュ フラクショヌマン エ デ ビオテクノロジーLaboratoire Francais du Fractionnement et des Biotechnologies | 病原体不活性化剤としてのグリコール |
AR087094A1 (es) | 2011-07-07 | 2014-02-12 | Gtc Biotherapeutics Inc | Formulaciones que estabilizan proteinas |
EP2556848A1 (en) * | 2011-08-08 | 2013-02-13 | Gambro Lundia AB | Separation material comprising saccharide ligands |
KR20140101331A (ko) | 2011-08-10 | 2014-08-19 | 라보라토이레 프란카이즈 듀 프락티온네먼트 에트 데스 바이오테크놀로지스 | 고도로 갈락토실화된 항체 |
US8859840B2 (en) * | 2011-09-08 | 2014-10-14 | Cynthia Katsingris | Multiple-use blood blotting devices for diabetics for use when monitoring blood glucose levels |
JP2015502370A (ja) | 2011-12-19 | 2015-01-22 | エルエフビー ユーエスエー インコーポレイテッドLfb Usa, Inc. | 炎症性疾患の治療のための組換えヒトα1−抗トリプシン |
TW201400499A (zh) | 2012-03-12 | 2014-01-01 | Revo Biolog Inc | 抗凝血酶用於治療妊娠毒血症之用途 |
MX2015001508A (es) | 2012-08-03 | 2015-04-08 | Lfb Usa Inc | El uso de antitrombina en la oxigenacion por membrana extracorporea. |
US20160039913A1 (en) | 2012-09-10 | 2016-02-11 | Lfb Usa, Inc. | The use of antibodies in treating hiv infection and suppressing hiv transmission |
KR20150145225A (ko) | 2013-02-13 | 2015-12-29 | 라보라토이레 프란카이즈 듀 프락티온네먼트 에트 데스 바이오테크놀로지스 | 변경된 글리코실화를 갖는 세툭시맙 및 이의 용도 |
EP2956485A2 (en) | 2013-02-13 | 2015-12-23 | Laboratoire Français du Fractionnement et des Biotechnologies | Highly galactosylated anti-her2 antibodies and uses thereof |
CN105308068A (zh) | 2013-02-13 | 2016-02-03 | 法国化学与生物科技实验室 | 高度半乳糖基化的抗TNF-α抗体及其用途 |
BR112015019348A2 (pt) | 2013-02-13 | 2017-08-22 | Lab Francais Du Fractionnement | Métodos para produção de proteína com glicosilação modificada e com sialilação aumentada, para aumentar a atividade de sialil transferase na glândula mamária e para produzir sialil transferase, proteína com glicosilação modificada ou proteína com sialilação aumentada, composição, sialil transferase, mamífero transgênico, e, célula epitelial mamária |
FR3005576B1 (fr) | 2013-05-15 | 2015-06-19 | Lab Francais Du Fractionnement | Procede d'inactivation d'une proteine prion |
PL3016729T3 (pl) | 2013-07-05 | 2020-09-07 | Laboratoire Français Du Fractionnement Et Des Biotechnologies Société Anonyme | Matryca do chromatografii powinowactwa |
FR3015484A1 (fr) | 2013-12-20 | 2015-06-26 | Lab Francais Du Fractionnement | Proteines recombinantes possedant une activite de facteur h |
WO2015100160A2 (en) | 2013-12-24 | 2015-07-02 | Lfb Usa, Inc. | Transgenic production of heparin |
CN106573973A (zh) | 2014-06-02 | 2017-04-19 | 法国化学与生物科技实验室 | Fc片段的产生 |
FR3022462B1 (fr) | 2014-06-18 | 2018-04-27 | Laboratoire Francais Du Fractionnement Et Des Biotechnologies | Composition orale d'anticorps anti-tnfalpha |
US20160158676A1 (en) | 2014-12-01 | 2016-06-09 | Laboratoire Français Du Fractionnement Et Des Biotechnologies | Method and apparatus for manipulating components of a filtration system |
CN107979973A (zh) | 2015-05-04 | 2018-05-01 | 法国化学与生物科技实验室 | Fc融合蛋白质的转基因生产 |
EP3307237A1 (en) | 2015-06-12 | 2018-04-18 | Laboratoire Français du Fractionnement et des Biotechnologies | Injectable composition of factor vii and fillers |
FR3038517B1 (fr) | 2015-07-06 | 2020-02-28 | Laboratoire Francais Du Fractionnement Et Des Biotechnologies | Utilisation de fragments fc modifies en immunotherapie |
-
2014
- 2014-07-04 PL PL14750552T patent/PL3016729T3/pl unknown
- 2014-07-04 ES ES14750552T patent/ES2793176T3/es active Active
- 2014-07-04 CA CA2916566A patent/CA2916566A1/fr not_active Abandoned
- 2014-07-04 AU AU2014285971A patent/AU2014285971A1/en not_active Abandoned
- 2014-07-04 EP EP14750552.3A patent/EP3016729B1/fr active Active
- 2014-07-04 WO PCT/FR2014/051734 patent/WO2015001277A1/fr active Application Filing
- 2014-07-04 JP JP2016522848A patent/JP2016532100A/ja not_active Ceased
- 2014-07-04 MX MX2016000063A patent/MX2016000063A/es unknown
- 2014-07-04 CN CN201480038362.8A patent/CN105358228A/zh active Pending
- 2014-07-04 KR KR1020167003199A patent/KR20160029840A/ko not_active Application Discontinuation
- 2014-07-04 TW TW103123198A patent/TW201509432A/zh unknown
- 2014-07-04 US US14/902,716 patent/US10611826B2/en active Active
-
2015
- 2015-12-30 IL IL243414A patent/IL243414A0/en unknown
Patent Citations (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US6069236A (en) * | 1993-06-14 | 2000-05-30 | Association Pour L'essor De La Transfusion Sanguine Dans La Region Du Nord | Immunoglobulin G concentrate for therapeutic use and process for producing said concentrate |
CN1196688A (zh) * | 1996-06-20 | 1998-10-21 | 巴克斯特国际有限公司 | 亲和性膜系统及其应用方法 |
US20040132979A1 (en) * | 2001-05-11 | 2004-07-08 | Chtourou Abdessatar Sami | Method for preparing human immunoglobulin concentrates for therapeutic use |
CN101098722A (zh) * | 2005-02-17 | 2008-01-02 | 弗雷森纽斯医疗护理德国有限责任公司 | 从液体尤其是血液中除去物质的装置 |
CN101346153A (zh) * | 2005-12-26 | 2009-01-14 | 分馏和生物工艺法国实验室公司 | 除去抗A、抗B抗体以及多反应性免疫球蛋白IgG的免疫球蛋白G(IgG)浓缩物 |
CN102921004A (zh) * | 2005-12-26 | 2013-02-13 | 分馏和生物工艺法国实验室公司 | 除去抗A、抗B抗体以及多反应性免疫球蛋白IgG的免疫球蛋白G(IgG)浓缩物 |
WO2013066251A1 (en) * | 2011-10-30 | 2013-05-10 | Glycorex Ab | Method for the reduction or elimination of one or more components from a blood product |
Cited By (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN108101981A (zh) * | 2018-01-15 | 2018-06-01 | 四川远大蜀阳药业股份有限公司 | 一种静注免疫球蛋白的生产工艺 |
CN108101981B (zh) * | 2018-01-15 | 2019-06-04 | 四川远大蜀阳药业有限责任公司 | 一种静注免疫球蛋白的生产工艺 |
WO2019137435A1 (zh) * | 2018-01-15 | 2019-07-18 | 四川远大蜀阳药业有限责任公司 | 一种静注免疫球蛋白的生产工艺 |
CN113166233A (zh) * | 2018-12-05 | 2021-07-23 | 西托索尔本茨公司 | 用于从人血浆和全血中去除抗a和/或抗b抗体的基于交联多糖的吸收剂 |
Also Published As
Publication number | Publication date |
---|---|
US20160168229A1 (en) | 2016-06-16 |
WO2015001277A1 (fr) | 2015-01-08 |
JP2016532100A (ja) | 2016-10-13 |
AU2014285971A1 (en) | 2016-02-04 |
TW201509432A (zh) | 2015-03-16 |
EP3016729B1 (fr) | 2020-03-25 |
IL243414A0 (en) | 2016-02-29 |
ES2793176T3 (es) | 2020-11-13 |
EP3016729A1 (fr) | 2016-05-11 |
US10611826B2 (en) | 2020-04-07 |
MX2016000063A (es) | 2016-03-01 |
CA2916566A1 (fr) | 2015-01-08 |
KR20160029840A (ko) | 2016-03-15 |
PL3016729T3 (pl) | 2020-09-07 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN105358228A (zh) | 亲和层析基质 | |
Kumar et al. | Affinity fractionation of lymphocytes using a monolithic cryogel | |
CN1327927C (zh) | 细胞分离组合物和方法 | |
KR101250095B1 (ko) | 항-A 및 항-B 항체 그리고 다중반응성 IgGs가 결핍된면역글로블린 G(IgG) 농축물 | |
RU2346039C2 (ru) | Композиция (варианты), способ и набор для разделения клеток | |
US10682640B2 (en) | Anionic exchange-hydrophobic mixed mode | |
EP1852443A1 (en) | Biocompatible three dimensional matrix for the immobilization of biological substances | |
CN105566442A (zh) | 一种降低单克隆抗体生产中宿主细胞蛋白含量的亲和纯化工艺 | |
FR3008097A1 (fr) | Matrice de chromatographie d'affinite | |
CN109964123A (zh) | 抗体的纯化方法 | |
Langley et al. | Another individual (JR) whose red blood cells appear to carry a hybrid MNSs sialoglycoprotein | |
FR3008098A1 (fr) | Matrice de chromatographie d'affinite a densite de ligands reduite | |
Ahmed et al. | Lymphoblastoid cell adhesion mediated by a dimeric and polymeric endogenous β‐galactoside‐binding lectin (galaptin) | |
WO1988007892A1 (en) | B lymphocyte separating material and body fluid clarifying material | |
US20180100023A1 (en) | Composition Enriched in Anti-A and /or Anti B Polyclonal Immunoglobulins | |
JP2022145698A (ja) | 精製方法 | |
CN117402833A (zh) | 分泌特异性抗体的杂交瘤细胞株、红细胞a抗体及应用 | |
BENACERRAF | DAVID H. KATZ, DIETER ARMERDING, MARTIN E. DORF, ZELIG ESHHAR | |
CN117362442A (zh) | 一种非对称双特异性抗体阴离子交换层析的洗脱方法 | |
JPS63252252A (ja) | Bリンパ球分離材、分離方法および分離器 | |
JP2020002079A (ja) | 分離剤の製造方法、分離剤の洗浄方法及び抗体の精製方法 | |
Kvarnfors et al. | ANTIBODY FORMATION IN CELL CULTURES: 2. The Effect of Rabbit Peritoneal Exudate Cells on Secondary in vitro IgG and IgM Antibody Responses to Poliovirus | |
CN104513303A (zh) | 用于筛选抗癌症医药组合物的多肽分子及其应用 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
C06 | Publication | ||
PB01 | Publication | ||
C10 | Entry into substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20160224 |
|
WD01 | Invention patent application deemed withdrawn after publication |