Characterization of anti-CD20 monoclonal antibody produced by transgenic silkworms (Bombyx mori).
In response to the successful use of monoclonal antibodies (mAbs) in the treatment of various diseases, systems for expressing recombinant mAbs using transgenic animals or plants have been widely developed. The silkworm (Bombyx mori) is a highly domesticated insect that has recently been used for the production of recombinant proteins. Because of their cost-effective breeding and relatively easy production scale-up, transgenic silkworms show great promise as a novel production system for mAbs. In this study, we established a transgenic silkworm stably expressing a human-mouse chimeric anti-CD20 mAb having the same amino acid sequence as rituximab, and compared its characteristics with rituximab produced by Chinese hamster ovary (CHO) cells (MabThera®). The anti-CD20 mAb produced in the transgenic silkworm showed a similar antigen-binding property, but stronger antibody-dependent cell-mediated cytotoxicity (ADCC) and weaker complement-dependent cytotoxicity (CDC) compared to MabThera. Post-translational modification analysis was performed by peptide mapping using liquid chromatography/mass spectrometry. There was a significant difference in the N-glycosylation profile between the CHO- and the silkworm-derived mAbs, but not in other post-translational modifications including oxidation and deamidation. The mass spectra of the N-glycosylated peptide revealed that the observed biological properties were attributable to the characteristic N-glycan structures of the anti-CD20 mAbs produced in the transgenic silkworms, i.e., the lack of the core-fucose and galactose at the non-reducing terminal. These results suggest that the transgenic silkworm may be a promising expression system for the tumor-targeting mAbs with higher ADCC activity.
a Division of Biological Chemistry and Biologicals; National Institute of Health Sciences ; Tokyo , Japan.
Minoru
M
Tada
b Transgenic Silkworm Research Unit; National Institute of Agrobiological Sciences ; Ibaraki , Japan.
Ken-ichiro
K
Tatematsu
a Division of Biological Chemistry and Biologicals; National Institute of Health Sciences ; Tokyo , Japan.
Akiko
A
Ishii-Watabe
a Division of Biological Chemistry and Biologicals; National Institute of Health Sciences ; Tokyo , Japan.
Akira
A
Harazono
a Division of Biological Chemistry and Biologicals; National Institute of Health Sciences ; Tokyo , Japan.|c Manufacturing Technology Research Association of Biologics ; Kobe , Japan.
Daisuke
D
Takakura
a Division of Biological Chemistry and Biologicals; National Institute of Health Sciences ; Tokyo , Japan.
Noritaka
N
Hashii
b Transgenic Silkworm Research Unit; National Institute of Agrobiological Sciences ; Ibaraki , Japan.
Hideki
H
Sezutsu
a Division of Biological Chemistry and Biologicals; National Institute of Health Sciences ; Tokyo , Japan.
Nana
N
Kawasaki
D000818:Animals; D030801:Animals, Genetically Modified; D000911:Antibodies, Monoclonal / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D000918:Antibody Specificity / Q000276:immunology*; D000920:Antibody-Dependent Cell Cytotoxicity / Q000276:immunology; D018951:Antigens, CD20 / Q000276:immunology*; D012831:Bombyx / Q000235:genetics*; D016466:CHO Cells; D045744:Cell Line, Tumor; D002853:Chromatography, Liquid; D003165:Complement System Proteins / Q000276:immunology; D006224:Cricetinae; D003412:Cricetulus; D003602:Cytotoxicity, Immunologic / Q000276:immunology; D006031:Glycosylation; D006801:Humans; D013058:Mass Spectrometry; D051379:Mice; D011994:Recombinant Proteins / Q000276:immunology* / Q000378:metabolism; D000069283:Rituximab / Q000235:genetics / Q000276:immunology / Q000378:metabolism
D000911:Antibodies, Monoclonal; D018951:Antigens, CD20; D011994:Recombinant Proteins; D000069283:Rituximab; D003165:Complement System Proteins
ADCC; N-glycosylation; monoclonal antibody; rituximab; transgenic silkworm