Preparation and separation of Fab and Fc fragments from human immunoglobulin G with papain digestion

TANG Siyuan; LIN Dongqiang; TONG Hongfei; YAO Shanjing

doi:10.3969/j.issn.0438-1157.2013.05.034

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Preparation and separation of Fab and Fc fragments from human immunoglobulin G with papain digestion

更新时间：2025-12-10

- Preparation and separation of Fab and Fc fragments from human immunoglobulin G with papain digestion
- CIESC Journal Issue 5, Pages: 1750-1756(2013)
- 作者机构：
  
  化学工程联合国家重点实验室, 浙江大学化学工程与生物工程学系,浙江,杭州,310027
- 作者简介：
- 基金信息：
- DOI：10.3969/j.issn.0438-1157.2013.05.034
  CLC： Q028
- Online First：05 May 2013，
  
  Published：05 May 2013
- 稿件说明：
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TANG Siyuan, LIN Dongqiang, TONG Hongfei, et al. Preparation and separation of Fab and Fc fragments from human immunoglobulin G with papain digestion[J]. CIESC Journal, 2013, (5): 1750-1756.
DOI：

TANG Siyuan, LIN Dongqiang, TONG Hongfei, et al. Preparation and separation of Fab and Fc fragments from human immunoglobulin G with papain digestion[J]. CIESC Journal, 2013, (5): 1750-1756. DOI： 10.3969/j.issn.0438-1157.2013.05.034.

摘要

为了制备抗体Fab和Fc片段

采用木瓜蛋白酶酶解人免疫球蛋白G(IgG)

通过优化酶解条件和色谱法分离过程

得到了纯度较高的Fab片段和Fc片段。考察了酶解pH、酶加入量、添加半胱氨酸和酶解时间等对IgG酶解过程的影响

优化了酶解条件

提高酶解效率

IgG转化率大于98%。酶解产物通过Protein A亲和色谱法和DEAE阴离子交换色谱法进行了纯化

分离得到Fc片段和Fab片段

收率分别为72.6%和40.1%。经SEC-HPLC分析

Fc片段纯度达95.7%

Fab片段纯度达96.6%。

Abstract

For the preparation of Fab and Fc fragments with high purity

human immunoglobulin G (hIgG) was treated with papain and its purification with chromatographic techniques was explored.The concentration of cystein

papain addition

pH and incubation time were optimized to improve digestion efficiency with papain

and conversion of IgG could reach more than 98%.Then Protein A affinity chromatography and DEAE anion-exchange chromatography were used to separate the Fab and Fc fragments from the digestion mixture of hIgG.Based on the SEC-HPLC analysis

the purity of Fab fragment was 96.6% with the yield of 40.1%

while the purity of Fc fragment was 95.7% with the yield of 72.6%.

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