公开(公告)号：US06462180B1

公开(公告)日：2002-10-08

申请号：US09449695

申请日：1999-11-24

Applicant: Wytold Lebing ,  Mark D. Chavez ,  David W. Ownby ,  Susan Trukawinski ,  Woody D. Wood

Inventor： Wytold Lebing ,  Mark D. Chavez ,  David W. Ownby ,  Susan Trukawinski ,  Woody D. Wood

IPC: C07K1700

CPC classification number: C07K14/8121 ,  Y10S530/831

Abstract: Purification of &agr;-1 proteinase inhibitor (&agr;-1 PI) from aqueous solutions, such as human plasma, is accomplished by precipitation of contaminating proteins from the aqueous solution, followed by dilution of the solution to adjust its conductivity, and passing of the solution through an anion exchange resin. The conductivity of the solution is adjusted so that the &agr;-1 PI binds to the anion exchange resin, while other contaminating proteins and solvent pass through the resin. Further purification may be accomplished by cation chromatography, which takes advantage of the fact that &agr;-1 PI does not bind to the cation column under certain conditions. Some embodiments of the invention also include virus removal and/or inactivation by methods such as nanofiltration and such as contact with a non-ionic detergent. The methods of the present invention result in greater yield, purity, and pathogenic clearance of plasma fractions than known methods.

Abstract translation: 从水溶液如人血浆中纯化α-1蛋白酶抑制剂（α-1 PI），通过从水溶液中沉淀污染蛋白质，然后稀释溶液以调节其导电性，并使溶液通过 通过阴离子交换树脂。 调节溶液的电导率使得α-1PI与阴离子交换树脂结合，而其它污染的蛋白质和溶剂通过树脂。 进一步纯化可以通过阳离子色谱来完成，其利用α-1PI在某些条件下不结合阳离子柱的事实。 本发明的一些实施方案还包括通过诸如纳滤的方法进行病毒去除和/或灭活，并且例如与非离子型洗涤剂的接触。 与已知方法相比，本发明的方法产生更高的产量，纯度和血浆级分的致病性清除。

公开(公告)号：US06307028B1

公开(公告)日：2001-10-23

申请号：US09270724

申请日：1999-03-17

Applicant: Wytold Lebing ,  Patricia Alred ,  Douglas C. Lee ,  Hanns-Ingolf Paul

Inventor： Wytold Lebing ,  Patricia Alred ,  Douglas C. Lee ,  Hanns-Ingolf Paul

IPC: C07K136

CPC classification number: A61L2/0088 ,  A61L2/18 ,  A61L2/23 ,  C07K16/065

Abstract: An improved process for the purification of antibodies from human plasma or other sources is disclosed. The process involves suspension of the antibodies at pH 3.8 to 4.5 followed by addition of caprylic acid and a pH shift to pH 5.0 to 5.2. A precipitate of contaminating proteins, lipids and caprylate forms and is removed, while the majority of the antibodies remain in solution. Sodium caprylate is again added to a final concentration of not less than about 15 mM. This solution is incubated for 1 hour at 25° C. to effect viral inactivation. A precipitate (mainly caprylate) is removed and the clear solution is diluted with purified water to reduce ionic strength. Anion exchange chromatography using two different resins is utilized to obtain an exceptionally pure IgG with subclass distribution similar to the starting distribution. The method maximizes yield and produces a gamma globulin with greater than 99% purity. The resin columns used to obtain a high yield of IgG, retain IgM and IgA, respectively. IgA and IgM may be eluted in high yield and purity.

Abstract translation: 公开了一种用于从人血浆或其他来源纯化抗体的改进方法。 该方法包括将抗体在pH 3.8至4.5下悬浮，然后加入辛酸，并将pH转变至pH5.5至5.2。 污染蛋白质，脂质和辛酸盐形式的沉淀物被除去，而大多数抗体保留在溶液中。 再加入辛酸钠，终浓度不低于约15mM。 将该溶液在25℃孵育1小时以进行病毒灭活。 除去沉淀物（主要是辛酸盐），并用净化水稀释澄清溶液以降低离子强度。 使用两种不同树脂的阴离子交换色谱法获得特异纯的IgG，其亚类分布类似于起始分布。 该方法使产量最大化，产生纯度高于99％的γ球蛋白。 用于获得高产量的IgG的树脂柱分别保留IgM和IgA。 可以以高产率和纯度洗脱IgA和IgM。

公开(公告)号：US09023797B2

公开(公告)日：2015-05-05

申请号：US13951070

申请日：2013-07-25

Applicant: Wytold Lebing ,  Doug Burns ,  Nathan Roth ,  Joann Hotta

Inventor： Wytold Lebing ,  Doug Burns ,  Nathan Roth ,  Joann Hotta

IPC: A61K38/38 ,  C07K14/76 ,  A01N37/02 ,  C12N7/00

CPC classification number: A01N37/02 ,  C07K14/76 ,  C12N7/00 ,  C12N2760/20263 ,  C12N2770/24363

Abstract: Described herein are methods for inactivating viruses using caprylate in solutions containing albumin.

Abstract translation: 本文描述了使用含有白蛋白的溶液中的辛酸盐灭活病毒的方法。

公开(公告)号：US20140163101A1

公开(公告)日：2014-06-12

申请号：US13951070

申请日：2013-07-25

Applicant: Wytold Lebing ,  Doug Burns ,  Nathan Roth ,  Joann Hotta

Inventor： Wytold Lebing ,  Doug Burns ,  Nathan Roth ,  Joann Hotta

IPC: A01N37/02 ,  C07K14/76

CPC classification number: A01N37/02 ,  C07K14/76 ,  C12N7/00 ,  C12N2760/20263 ,  C12N2770/24363

Abstract: Described herein are methods for inactivating viruses using caprylate in solutions containing albumin.

Abstract translation: 本文描述了使用含有白蛋白的溶液中的辛酸酯灭活病毒的方法。

公开(公告)号：US20110237781A1

公开(公告)日：2011-09-29

申请号：US13002429

申请日：2009-07-17

Applicant: Wytold Lebing ,  Scott A. Cook ,  Christopher A. Dadd

Inventor： Wytold Lebing ,  Scott A. Cook ,  Christopher A. Dadd

IPC: C07K1/36

CPC classification number: C07K14/8125

Abstract: Purification of α-1 proteinase inhibitor (α-1 PI) from solutions comprising α-1 PI is accomplished using hydrophobic interaction chromatography (HIC). In some embodiments, purification of α-1 PI is accomplished by precipitation of contaminating proteins from a starting solution comprising α-1 PI, such as human plasma, followed by anion exchange resin chromatography prior to HIC. Further purification may be accomplished by an optional cation exchange chromatography subsequent to anion exchange chromatography but prior to HIC. Some embodiments of the invention also include virus removal and/or inactivation by methods such as nano filtration and such as contact with a non-ionic detergent. The methods of the present invention result in greater yield, purity, and pathogenic clearance of plasma fractions than known methods.

Abstract translation: 使用疏水相互作用色谱（HIC）实现从包含α-1 PI的溶液中纯化α-1蛋白酶抑制剂（α-1 PI）。 在一些实施方案中，通过从包含α-1PI的起始溶液（例如人血浆）中沉淀污染性蛋白，然后在HIC之前进行阴离子交换树脂色谱来完成α-1 PI的纯化。 进一步纯化可以通过阴离子交换层析之后但在HIC之前的任选的阳离子交换层析来完成。 本发明的一些实施方案还包括通过诸如纳米过滤的方法和诸如与非离子型洗涤剂接触的病毒去除和/或灭活。 与已知方法相比，本发明的方法产生更高的产量，纯度和血浆级分的致病性清除。