Purification of monoclonal antibody against Ebola GP1 protein expressed in Nicotiana benthamiana.
Monoclonal antibodies (mAbs) are one of the fastest growing drug molecules targeting the treatment of diseases ranging from arthritis, immune disorders, and infectious diseases to cancer. Due to its unique application principle, antibodies are commonly produced in large quantities. Plants, such as Nicotiana benthamiana, offer a unique production platform for bio-therapeutics due to their ability to produce large amounts of biomolecules in a relatively quick manner. However, purification of a target protein from plant is an arduous task due to the presence of toxic compounds in ground plant tissue and the large quantities of plant tissues to be processed. Here, a process was developed prior to the chromatographic purification of a mAb against Ebola GP1 protein expressed in N. benthamiana. The process includes a diafiltration step and a charged polyelectrolyte precipitation. The diafiltration step significantly improved the precipitation efficiency, reducing the usage of polyelectrolyte by more than 2000 fold while improving the native plant protein removed from 60% to 80%. The mAb can then be purified to near homogeneity judging from SDS-PAGE by either Protein A affinity chromatography or a tandem of hydrophobic interaction chromatography and a hydrophobic charge induction chromatography. The purified mAbs were shown to retain their binding specificity to irradiated Ebola virus.
Department of Biological Systems Engineering, Virginia Tech, Blacksburg, VA 24061, USA.
Andrew
A
Fulton
The Biodesign Institute and School of Life Sciences, Arizona State University, Tempe, AZ, USA.
Huafang
H
Lai
The Biodesign Institute and School of Life Sciences, Arizona State University, Tempe, AZ, USA.
Qiang
Q
Chen
Department of Biological Systems Engineering, Virginia Tech, Blacksburg, VA 24061, USA. Electronic address: chzhang2@vt.edu.
Chenming
C
Zhang
D000911:Antibodies, Monoclonal / Q000302:isolation & purification*; D000914:Antibodies, Viral / Q000302:isolation & purification*; D029043:Ebolavirus / Q000235:genetics*; D004591:Electrophoresis, Polyacrylamide Gel; D015870:Gene Expression; D011485:Protein Binding; D014026:Nicotiana / Q000235:genetics*; D014759:Viral Envelope Proteins / Q000235:genetics* / Q000378:metabolism
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D014759:Viral Envelope Proteins; C113189:envelope glycoprotein, Ebola virus
Antibody purification; Ebola virus; Monoclonal antibody; Nicotiana benthamiana; Transgenic plant; Transient expression