Considerations for extraction of monoclonal antibodies targeted to different subcellular compartments in transgenic tobacco plants

Summary Monoclonal antibody production from transgenic tobacco plants offers many advantages over other heterologous production systems, creating the prospect of production at a scale that will allow new prophylactic and therapeutic applications in global human and animal health. However, informatio...

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Bibliographic Details
Published in:Plant biotechnology journal 2008-09, Vol.6 (7), p.733-748
Main Authors: Hassan, Sally, Van Dolleweerd, Craig J., Ioakeimidis, Fotis, Keshavarz-Moore, Eli, Ma, Julian K.-C.
Format: Article
Language:English
Subjects:
Antibodies, Monoclonal - genetics
Antibodies, Monoclonal - isolation & purification
Antibodies, Monoclonal - metabolism
Biological and medical sciences
Biotechnology
Biotechnology - methods
Cell Compartmentation
Cell Membrane - chemistry
Chemical Fractionation - methods
Endoplasmic Reticulum - chemistry
extraction
Fundamental and applied biological sciences. Psychology
Glycosylation
Hydrogen-Ion Concentration
leaves
monoclonal antibody
Nicotiana - chemistry
Nicotiana - genetics
Plant Leaves - chemistry
Plant Proteins - genetics
Plant Proteins - isolation & purification
Plants, Genetically Modified - chemistry
Plants, Genetically Modified - metabolism
Protein Transport
Recombinant Fusion Proteins - isolation & purification
Recombinant Fusion Proteins - metabolism
Reproducibility of Results
Temperature
tobacco
transgenic
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Summary:Summary Monoclonal antibody production from transgenic tobacco plants offers many advantages over other heterologous production systems, creating the prospect of production at a scale that will allow new prophylactic and therapeutic applications in global human and animal health. However, information on the major processing factors to consider for large‐scale purification of antibodies from transgenic plants is currently limited, and is in urgent need of attention. The purpose of this project was to investigate methods for the initial extraction of recombinant immunoglobulin G (IgG) antibodies from transgenic tobacco leaf tissue. Three different transgenic plant lines were studied in order to establish the parameters for optimal extraction of monoclonal antibodies that accumulate in the apoplasm, at the plasma membrane or within the endoplasmic reticulum. For each transgenic line, seven techniques for physical extraction were compared. The factors that determine the optimal extraction of antibodies from plants have a direct influence on the initial choice of expression strategy, and so must be considered at an early stage. The use of small‐scale techniques that are applicable to large‐scale purification was a particularly important consideration. The optimal extraction technique varied with the target location of IgG in the plant cell, and the dependence of antibody yield on the physical extraction methodology employed, the pH of the extraction buffer and the extraction temperature was demonstrated in each case. The addition of detergent to the extraction buffer may improve the yield, but this was found to be dependent on the site of accumulation of IgG within the plant cell.
ISSN:1467-7644
1467-7652
DOI:10.1111/j.1467-7652.2008.00354.x