PMID- 18793269
OWN - NLM
STAT- MEDLINE
DCOM- 20090130
LR  - 20231213
IS  - 1467-7652 (Electronic)
IS  - 1467-7644 (Linking)
VI  - 7
IP  - 1
DP  - 2009 Jan
TI  - Plant pharming of a full-sized, tumour-targeting antibody using different 
      expression strategies.
PG  - 59-72
LID - 10.1111/j.1467-7652.2008.00371.x [doi]
AB  - The aims of this work were to obtain a human antibody against the 
      tumour-associated antigen tenascin-C (TNC) and to compare the yield and quality 
      of plant-produced antibody in either stable transgenics or using a transient 
      expression system. To this end, the characterization of a full-sized human 
      immunoglobulin G (IgG) [monoclonal antibody H10 (mAb H10)], derived from a 
      selected single-chain variable fragment (scFv) and produced in plants, is 
      presented. The human mAb gene was engineered for plant expression, and Nicotiana 
      tabacum transgenic lines expressing both heavy (HC) and light (LC) chain were 
      obtained and evaluated for antibody expression levels, in vivo assembly and 
      functionality. Affinity-purified H10 from transgenics (yield, 0.6-1.1 mg/kg fresh 
      weight) revealed that more than 90% of HC was specifically degraded, leading to 
      the formation of functional antigen-binding fragments (Fab). Consequently, H10 
      was transiently expressed in Nicotiana benthamiana plants through an 
      Agrobacterium-mediated gene-transfer system. Moreover, the use of the p19 
      silencing suppressor gene from artichoke mottled crinkle virus raised antibody 
      expression levels by an order of magnitude (yields of purified H10, 50-100 mg/kg 
      fresh weight). Approximately 75% of purified protein consisted of full-sized 
      antibody functionally binding to TNC (K(D) = 14 nm), and immunohistochemical 
      analysis on tumour tissues revealed specific accumulation around tumour blood 
      vessels. The data indicate that the purification yields of mAb H10, using a 
      transient expression system boosted by the p19 silencing suppressor, are 
      exceptionally high when compared with the results reported previously, providing 
      a technique for the over-expression of anticancer mAbs by a rapid, 
      cost-effective, molecular farming approach.
FAU - Villani, Maria Elena
AU  - Villani ME
AD  - ENEA, Dipartimento BAS, Sezione Genetica e Genomica Vegetale, C.R. Casaccia, Via 
      Anguillarese 301, I-00123, Rome, Italy.
FAU - Morgun, Bogdan
AU  - Morgun B
FAU - Brunetti, Patrizia
AU  - Brunetti P
FAU - Marusic, Carla
AU  - Marusic C
FAU - Lombardi, Raffaele
AU  - Lombardi R
FAU - Pisoni, Ivan
AU  - Pisoni I
FAU - Bacci, Camilla
AU  - Bacci C
FAU - Desiderio, Angiola
AU  - Desiderio A
FAU - Benvenuto, Eugenio
AU  - Benvenuto E
FAU - Donini, Marcello
AU  - Donini M
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20080910
PL  - England
TA  - Plant Biotechnol J
JT  - Plant biotechnology journal
JID - 101201889
RN  - 0 (Antibodies, Monoclonal)
RN  - 0 (Antibodies, Neoplasm)
RN  - 0 (Immunoglobulin Variable Region)
RN  - 0 (Recombinant Proteins)
RN  - 0 (Tenascin)
SB  - IM
MH  - Amino Acid Sequence
MH  - Animals
MH  - Antibodies, Monoclonal/*biosynthesis/genetics/immunology
MH  - Antibodies, Neoplasm/*biosynthesis/genetics/immunology
MH  - Gene Expression
MH  - Humans
MH  - Immunoglobulin Variable Region/biosynthesis/genetics/immunology
MH  - Mice
MH  - Molecular Sequence Data
MH  - Neoplasms, Experimental/immunology
MH  - Plants, Genetically Modified/genetics/immunology/*metabolism
MH  - Protein Engineering
MH  - Recombinant Proteins/biosynthesis/genetics/immunology
MH  - Tenascin/*antagonists & inhibitors
MH  - Nicotiana/genetics/metabolism
MH  - Transformation, Genetic
EDAT- 2008/09/17 09:00
MHDA- 2009/01/31 09:00
CRDT- 2008/09/17 09:00
PHST- 2008/09/17 09:00 [pubmed]
PHST- 2009/01/31 09:00 [medline]
PHST- 2008/09/17 09:00 [entrez]
AID - PBI371 [pii]
AID - 10.1111/j.1467-7652.2008.00371.x [doi]
PST - ppublish
SO  - Plant Biotechnol J. 2009 Jan;7(1):59-72. doi: 10.1111/j.1467-7652.2008.00371.x. 
      Epub 2008 Sep 10.