公开(公告)号：US06740740B2

公开(公告)日：2004-05-25

申请号：US09962527

申请日：2001-09-24

Applicant: Stephen J. Garger ,  R. Barry Holtz ,  Michael J. McCulloch ,  Thomas H. Turpen

Inventor： Stephen J. Garger ,  R. Barry Holtz ,  Michael J. McCulloch ,  Thomas H. Turpen

IPC: C07K136

CPC classification number: C12N15/8258 ,  C07K14/415 ,  C07K16/00 ,  C07K16/16 ,  C07K2317/13 ,  C07K2319/00 ,  C12N7/00 ,  C12N15/8203 ,  C12N15/8242 ,  C12N15/8257 ,  C12N2710/00051 ,  C12N2770/00051 ,  Y10S977/904

Abstract: The present invention features a method for isolating and purifying proteins and peptides of interest from a plant host, which is applicable on a larger scale. Moreover, the present invention provides a more efficient method for isolating proteins and peptides of interest than those methods described in the prior art. In general, the present method of isolating proteins and peptides of interest comprises the steps of homogenizing a plant to produce a green juice, adjusting the pH of and heating the green juice, separating the target protein/peptide from other components of the green juice by one or more cycles of centrifugation and/or resuspension, and finally purifying proteins and peptides by such procedures as chromatography and/or salt precipitation.

Abstract translation: 本发明特征在于从植物宿主中分离和纯化目的蛋白质和肽的方法，该方法可适用于较大规模。 此外，本发明提供了一种比现有技术中描述的方法更有效的分离蛋白质和肽的方法。 通常，本发明的分离蛋白质和肽的方法包括以下步骤：将植物均质化以产生绿汁，调节绿汁的pH并加热绿汁，将目标蛋白质/肽与绿汁的其它成分分离，通过 离心和/或再悬浮的一个或多个循环，并且最后通过诸如色谱法和/或盐沉淀的方法纯化蛋白质和肽。

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。