Large-scale purification of an antibody directed against hepatitis B surface antigen from transgenic tobacco plants

被引：59

作者：

Valdés, R ^{[1
]}

Gómez, L ^{[1
]}

Padilla, S ^{[1
]}

Brito, J ^{[1
]}

Reyes, B ^{[1
]}

Alvarez, T ^{[1
]}

Mendoza, O ^{[1
]}

Herrera, O ^{[1
]}

Ferro, W ^{[1
]}

Pujol, M ^{[1
]}

Leal, V ^{[1
]}

Linares, M ^{[1
]}

Hevia, Y ^{[1
]}

García, C ^{[1
]}

Milá, L ^{[1
]}

García, O ^{[1
]}

Sánchez, R ^{[1
]}

Acosta, A ^{[1
]}

Geada, D ^{[1
]}

Paez, R ^{[1
]}

Vega, JL ^{[1
]}

Borroto, C ^{[1
]}

机构：

[1] Plant Derived Antibody Prod & Generat Grp, Ctr Genet Engn & Biotechnol, Havana 10600, Cuba

来源：

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS | 2003年 / 308卷 / 01期

关键词：

plantibody; monoclonal antibody; transgenic plant; expanded bed adsorption; tobacco;

D O I：

10.1016/S0006-291X(03)01335-4

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

The application of bioengineering to plants for production of biological products for human and animal use has expanded in recent years. The reasons for this expansion are several and include advances in the technology for novel production systems and the need for very large quantities of therapeutic proteins. The process of growing pharmaceutical proteins in plants, extracting, and purifying is a hard task considering the lack of available information concerning these topics. In this work, a recombinant murine monoclonal antibody specific for the hepatitis B surface antigen, expressed in stably transformed transgenic Nicotiana tabacum plants, was purified by means of a recombinant protein A Streamline chromatography as the main purification step. The antibody expression level varied with the age of the plants and the number of harvests from 40 to 15 mug/ml and the maximum process yield was about 25 mg of plantibody/kg of biomass. Protein A Streamline chromatography was successfully used in the purification process yielding a recovery of about 60% and a plantibody SDS-PAGE purity of over 90% but unexpectedly, previous clarification steps could not be totally avoided. The amino acid sequence recognized by this affinity purified plantibody was similar to its murine counterpart verifying the potentiality of plants to replace animals or bioreactors for large-scale production of this monoclonal antibody. (C) 2003 Published by Elsevier Inc.

引用

页码：94 / 100

页数：7

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