公开(公告)号：US06946250B2

公开(公告)日：2005-09-20

申请号：US10000526

申请日：2001-12-04

Applicant: Helge Bastian ,  Simone Gauch ,  Metin Colpan ,  Petra Feuser

Inventor： Helge Bastian ,  Simone Gauch ,  Metin Colpan ,  Petra Feuser

IPC: C07H1/06 ,  B01J20/281 ,  B01J20/283 ,  B01J20/285 ,  C07H1/08 ,  C07H21/00 ,  C07H21/02 ,  C07H21/04 ,  C12N15/09 ,  C12N15/10 ,  C12Q1/68 ,  G01N30/26 ,  G01N30/88 ,  C12P19/34

CPC classification number: C12N15/1006 ,  C12N15/101 ,  C12Q1/6806

Abstract: The present invention pertains to a process for the chromatographic separation of nucleic acid mixtures into their double-stranded and single-stranded nucleic acid fractions by simultaneously absorbing said nucleic acids as a whole to a mineral support, followed by separation into double-stranded and single-stranded nucleic acids by fractional elution, or by selectively absorbing double-stranded or single-stranded nucleic acid of a liquid sample to a mineral support, as well as solutions and a kit for performing the process according to the invention.

Abstract translation: 本发明涉及通过将所述核酸整体同时吸收到矿物载体上，然后将其分离成双链和单链核酸的单链和单链核酸部分，将核酸混合物色谱分离成双链和单链核酸部分 通过分批洗脱或通过选择性地将液体样品的双链或单链核酸吸收到矿物载体，以及用于执行根据本发明的方法的溶液和试剂盒来代替。

公开(公告)号：US07510826B2

公开(公告)日：2009-03-31

申请号：US11412130

申请日：2006-04-27

Applicant: Metin Colpan ,  Peter Moritz ,  Joachim Schorr

Inventor： Metin Colpan ,  Peter Moritz ,  Joachim Schorr

IPC: C12Q1/00 ,  G01N30/02 ,  G01N31/00 ,  G01N33/48

CPC classification number: B01D15/363 ,  B01D15/3804 ,  C12N15/101 ,  C12N15/87 ,  Y10S435/81 ,  Y10S435/82 ,  Y10S436/808 ,  Y10T436/10 ,  G01N30/461

Abstract: A process for the depletion or removal of endotoxins from preparations containing active ingredients designated for therapeutical use which are obtained from natural sources by genetic engineering and/or biotechnology by treatment with chromatographic material wherein said natural sources are lysed, the fractions obtained are optionally centrifuged, filtrated or treated with affinity chromatographic methods; said fractions are preincubated with an aqueous salt solution and detergents, treated with anion exchange material and then washed with another salt solution, and the active ingredients are eluted from the anion exchanger, followed by further purification in a per se known manner.

Abstract translation: 从含有指定用于治疗用途的活性成分的制剂中消耗或除去内毒素的方法，其通过遗传工程和/或生物技术从天然来源获得，其中所述天然来源被裂解，所得的级分任选地离心， 用亲和层析法过滤或处理; 所述级分用盐水溶液和洗涤剂预先加热，用阴离子交换材料处理，然后用另一种盐溶液洗涤，活性成分从阴离子交换剂中洗脱，然后以本身已知的方式进一步纯化。

公开(公告)号：US06297371B1

公开(公告)日：2001-10-02

申请号：US09253702

申请日：1999-02-22

Applicant: Metin Colpan ,  Joachim Schorr ,  Peter Moritz

Inventor： Metin Colpan ,  Joachim Schorr ,  Peter Moritz

IPC: C07H2100

CPC classification number: B01D15/363 ,  B01D15/3804 ,  C12N15/101 ,  C12N15/87 ,  Y10S435/81 ,  Y10S435/82 ,  Y10S436/808 ,  Y10T436/10 ,  G01N30/461

Abstract: A process for the isolation and purification of nucleic acids and/or oligonucleotides for use in gene therapy wherein said nucleic acids and/or oligonucleotides are isolated or purified from an essentially biological source, characterized in that said essentially biological sources are lysed, the fractions obtained are optionally freed or depleted from the remainder of said biological sources by per se known mechanical methods, such as centrifugation, filtration; the fractions thus treated are subsequently treated with affinity chromatographic material or with inorganic chromatographic material for the removal of endotoxins; followed by isolation of said nucleic acids and/or oligonucleotides on an anion exchanger which is designed such that DNA begins to desorb from the anion exchanger only at an ionic strength corresponding to a sodium chloride solution of a concentration higher by at least 100 mM than one corresponding to the ionic strength at which RNA begins to desorb from the anion exchanger material.

Abstract translation: 用于分离和纯化用于基因治疗的核酸和/或寡核苷酸的方法，其中所述核酸和/或寡核苷酸从基本上生物来源分离或纯化，其特征在于基本上生物来源被裂解，获得的级分为 通过本身已知的机械方法，例如离心，过滤，任选地从所述生物源的剩余部分中释放或消耗;随后用亲和层析材料或用无机色谱材料处理如此处理的级分以除去内毒素; 然后在阴离子交换器上分离所述核酸和/或寡核苷酸，所述阴离子交换器被设计为使得DNA开始从阴离子交换剂解吸离子，其离子强度对应于浓度高于一个相应浓度至少100mM的氯化钠溶液 到RNA开始从阴离子交换材料解吸的离子强度。

公开(公告)号：US06277648B1

公开(公告)日：2001-08-21

申请号：US08253152

申请日：1994-06-02

Applicant: Metin Colpan

Inventor： Metin Colpan

IPC: B01D3700

CPC classification number: C12N15/101 ,  B01D39/00 ,  C12N15/1017 ,  Y10T436/25375 ,  Y10T436/255

Abstract: Described is a method of isolating cell components, such as nucleic acids, from natural sources by filtering a sample of the digested natural sources such as cells or cell fragments. The method is characterized in that the sample is passed through a filter, the pore size of which decreases in the direction of flow of the sample through the filter.

Abstract translation: 描述了通过过滤消化的天然来源（例如细胞或细胞碎片）的样品从天然来源分离细胞成分（例如核酸）的方法。 该方法的特征在于，样品通过过滤器，其孔径在样品通过过滤器的流动方向上减小。

公开(公告)号：US06274371B1

公开(公告)日：2001-08-14

申请号：US08809072

申请日：1997-06-19

Applicant: Metin Colpan

Inventor： Metin Colpan

IPC: C12M133

CPC classification number: C12N15/101 ,  C12N15/87 ,  Y10S435/975

Abstract: A filtration process for the preparation of nucleic acids from natural sources is disclosed. The sources containing nucleic acids are lysed; the lysate is left to rest for some time; the resulting lysate passes a filter layer. The filter layer is selected from silica gel, aluminum oxide or packed diatomaceous earth, or interwoven or cemented non-wovens of glass fibers and silica gel. Other filter layers include cellulose, paper, compressed paper, and paper non-wovens. The fraction leaving the filter layer is then collected and the nucleic acid is subsequently isolated and purified from the collected fraction. Furthermore, the filter layer has not been modified with anion exchange groups.

Abstract translation: 公开了一种用于从天然来源制备核酸的过滤方法。 含有核酸的来源被裂解; 裂解物待休息一段时间; 所得裂解物通过过滤层。 过滤层选自硅胶，氧化铝或填充的硅藻土，或玻璃纤维和硅胶的交织或粘合无纺布。 其它过滤层包括纤维素，纸，压缩纸和纸无纺布。 然后收集离开过滤层的级分，随后从收集的级分中分离和纯化核酸。 此外，过滤层未用阴离子交换基团修饰。

公开(公告)号：US5990301A

公开(公告)日：1999-11-23

申请号：US687529

申请日：1996-10-18

Applicant: Metin Colpan ,  Joachim Schorr ,  Peter Moritz

Inventor： Metin Colpan ,  Joachim Schorr ,  Peter Moritz

IPC: C12N15/10 ,  C12N15/87 ,  C07H21/00 ,  C12N1/08

CPC classification number: C12N15/87 ,  C12N15/101 ,  Y10S435/81

Abstract: A process for the isolation and purification of nucleic acids and/or oligonucleotides for use in gene therapy wherein nucleic acids and/or oligonucleotides are isolated or purified from an essentially biological source, characterized in thatessentially biological sources are lysed, the fractions obtained are optionally freed or depleted from the remainder of biological sources by per se known mechanical methods, such as centrifugation, filtration;the fractions thus treated are subsequently treated with affinity chromatographic material or with inorganic chromatographic material for the removal of endotoxins; followed byisolation of nucleic acids and/or oligonucleotides on an anion exchanger which is designed such that DNA begins to desorb from the anion exchanger only at an ionic strength corresponding to a sodium chloride solution of a concentration higher by at least 100 mM than one corresponding to the ionic strength at which RNA begins to desorb from the anion exchanger material.

Abstract translation: PCT No.PCT / EP95 / 00389 371日期：1996年10月18日 102（e）日期1996年10月18日PCT提交1995年2月3日PCT公布。 第WO95 / 21177号公报 日期1995年8月10日一种用于分离和纯化用于基因治疗的核酸和/或寡核苷酸的方法，其中从基本上生物来源分离或纯化核酸和/或寡核苷酸，其特征在于基本上生物来源被裂解， 通过本身已知的机械方法，例如离心，过滤，任选地从所述生物来源的其余部分中释放或消耗所得的级分。 随后用亲和层析材料或用无机色谱材料处理如此处理的级分以除去内毒素; 随后在阴离子交换器上分离核酸和/或寡核苷酸，其被设计为使得DNA开始从阴离子交换剂解吸，仅在对应于浓度高于一个浓度的至少100mM的氯化钠溶液的离子强度 到RNA开始从阴离子交换材料解吸的离子强度。

公开(公告)号：US08247545B1

公开(公告)日：2012-08-21

申请号：US08796040

申请日：1997-02-05

Applicant: Metin Colpan

Inventor： Metin Colpan

IPC: C07H21/00

CPC classification number: C12N15/101 ,  B01D39/00 ,  C12N15/1017 ,  Y10T436/25375 ,  Y10T436/255

Abstract: There is described is a process for the isolation and purification of nucleic acids such as plasmid or genomic DNA from cells or other sources, wherein a) the cells containing nucleic acids are digested and cell debris is removed, or other samples containing nucleic acids are treated with anion exchangers, namely, in buffer solutions of low ionic strength, b) thereafter, the nucleic acids are desorbed from the anion exchanger using a buffer of high ionic strength, in order to be subsequently c) treated in said buffer of high ionic strength or in the presence of lower alcohols and/or poly(ethylene glycol) with a mineral support material, with adsorption of the nucleic acid to the surface of the mineral support materials, whereupon d) desorption of the nucleic acid is effected using water or a buffer solution of low ionic strength. The device for operating the inventive process consists of a hollow body (1) with an inlet opening (7) and an outlet opening (8), wherein in the hollow body (1), between two securing means (5, 6), a powdered first material based on silica gel (10) is arranged, and a second material (11) is placed between the first material (10) and the outlet opening (8), the first and second materials (10, 11) having different adsorption characteristics for nucleic acids.

Abstract translation: 描述了用于分离和纯化来自细胞或其他来源的核酸如质粒或基因组DNA的方法，其中a）将含有核酸的细胞消化并除去细胞碎片，或处理含有核酸的其它样品 具有阴离子交换剂，即在低离子强度的缓冲溶液中，b）此后，使用高离子强度的缓冲液将核酸从阴离子交换剂中解吸，以便随后c）在所述高离子强度缓冲液中处理 或在具有矿物载体材料的低级醇和/或聚（乙二醇）的存在下，将核酸吸附到矿物载体材料的表面，因此d）核酸的解吸是使用水或 缓冲溶液的低离子强度。 用于操作本发明方法的装置包括具有入口开口（7）和出口开口（8）的空心体（1），其中在中空体（1）中，在两个固定装置（5,6）之间， 布置基于硅胶（10）的粉末状第一材料，并且第二材料（11）被放置在第一材料（10）和出口（8）之间，第一和第二材料（10,11）具有不同的吸附 核酸的特征。

公开(公告)号：US07109322B2

公开(公告)日：2006-09-19

申请号：US10254845

申请日：2002-09-26

Applicant: Metin Colpan ,  Peter Moritz ,  Joachim Schorr

Inventor： Metin Colpan ,  Peter Moritz ,  Joachim Schorr

IPC: C07H21/00 ,  C12N1/00 ,  C12N1/08 ,  C12P19/34

CPC classification number: B01D15/363 ,  B01D15/3804 ,  C12N15/101 ,  C12N15/87 ,  Y10S435/81 ,  Y10S435/82 ,  Y10S436/808 ,  Y10T436/10 ,  G01N30/461

Abstract: A process for the depletion or removal of endotoxins from preparations containing active ingredients designated for therapeutical use which are obtained from natural sources by genetic engineering and/or biotechnology by treatment with chromatographic material wherein said natural source are lysed, the fractions obtained are optionally centrifuged, filtrated or treated with affinity chromatographic methods; said fractions are preincubated with an aqueous salt solution and detergents, treated with anion exchange material and then washed with another salt solution, and the active ingredients are eluted from the anion exchanger, followed by further purification in a per se known manner.

公开(公告)号：US5652141A

公开(公告)日：1997-07-29

申请号：US442100

申请日：1995-05-16

Applicant: Karsten Henco ,  Metin Colpan ,  Petra Feuser

Inventor： Karsten Henco ,  Metin Colpan ,  Petra Feuser

IPC: C12N15/10 ,  C12Q1/68 ,  C12N1/08 ,  C07H1/06

CPC classification number: C12N15/101 ,  C12N15/1006 ,  C12Q1/6806

Abstract: A device and a process for isolating nucleic acids by lysing intact cells and removing nucleic acids emerging from the lysed cells by the following steps:a) the cells are immobilized in a porous matrix, with the size of matrix voids being in the range of the type of cell to be lysed;b) the cells are lysed;c) the nucleic acids are fixated on the matrix surface, and subsequentlyd) are eluted.

Abstract translation: 通过以下步骤裂解完整细胞和从裂解的细胞中除去出现的核酸来分离核酸的装置和方法：a）将细胞固定在多孔基质中，其中基质空隙的大小在 待裂解的细胞类型; b）细胞裂解; c）将核酸固定在基质表面上，随后d）被洗脱。

公开(公告)号：US4699717A

公开(公告)日：1987-10-13

申请号：US830708

申请日：1986-02-14

Applicant: Detlev Riesner ,  Metin Colpan

Inventor： Detlev Riesner ,  Metin Colpan

IPC: B01D15/08 ,  B01J20/28 ,  B01J20/32

CPC classification number: B01D15/08 ,  B01J20/103 ,  B01J20/28004 ,  B01J20/28057 ,  B01J20/28085 ,  B01J20/28095 ,  B01J20/286 ,  B01J20/3204 ,  B01J20/3219 ,  B01J20/3257 ,  B01J20/3259 ,  B01J20/3263 ,  B01D15/34 ,  B01D15/362 ,  B01D15/363 ,  B01J2220/54 ,  B01J2220/58

Abstract: A process for the chromatographic separation of nucleic acid using a chromatographic carrier material is described in which the surface of the carrier material is specially modified.

Abstract translation: 描述了使用色谱载体材料色谱分离核酸的方法，其中载体材料的表面被特别改性。