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1.
《MABS-AUSTIN》2013,5(3):230-236
ADCC, antibody-dependent cellular cytotoxicity; CDC, complement-dependent cytotoxicity; Fc, antibody constant region; FcγRIIIa, human Fcγ-receptor IIIa; IgG, immunoglobulin G; NK cell, natural killer cell; CHO, Chinese hamster ovary; EPO, erythropoietin; Glc, glucose; Man, mannose; GlcNAc, N-acetylglucosamine; Gal, galactose; NANA; N-acetylneuraminic acid; FUT8, α-1,6 fucosyltransferase; GMD, GDP-mannose 4,6-dehydratase; FX, GDP-keto-6-deoxymannose 3,5-epimerase/4-reductase; GFT, GDP-fucose transporter; siRNA, short interfering RNA; GnTIII, β-1,4-N-acetylglucosaminyltransferase; ManII, α-mannosidase II 相似文献
2.
Non-fucosylated therapeutic antibodies: the next generation of therapeutic antibodies 总被引:1,自引:0,他引:1
Katsuhiro Mori Shigeru Iida Naoko Yamane-Ohnuki Yutaka Kanda Reiko Kuni-Kamochi Ryosuke Nakano Harue Imai-Nishiya Akira Okazaki Toyohide Shinkawa Akihito Natsume Rinpei Niwa Kenya Shitara Mitsuo Satoh 《Cytotechnology》2007,55(2-3):109-114
Therapeutic antibody IgG1 has two N-linked oligosaccharide chains bound to the Fc region. The oligosaccharides are of the complex biantennary type, composed of a trimannosyl core structure with the presence or absence of core fucose, bisecting N-acetylglucosamine (GlcNAc), galactose, and terminal sialic acid, which gives rise to structural heterogeneity. Both human serum IgG and therapeutic antibodies are well known to be heavily fucosylated. Recently, antibody-dependent cellular cytotoxicity (ADCC), a lytic attack on antibody-targeted cells, has been found to be one of the critical effector functions responsible for the clinical efficacy of therapeutic antibodies such as anti-CD20 IgG1 rituximab (Rituxan®) and anti-Her2/neu IgG1 trastuzumab (Herceptin®). ADCC is triggered upon the binding of lymphocyte receptors (FcγRs) to the antibody Fc region. The activity is dependent on the amount of fucose attached to the innermost GlcNAc of N-linked Fc oligosaccharide via an α-1,6-linkage, and is dramatically enhanced by a reduction in fucose. Non-fucosylated therapeutic antibodies show more potent efficacy than their fucosylated counterparts both in vitro and in vivo, and are not likely to be immunogenic because their carbohydrate structures are a normal component of natural human serum IgG. Thus, the application of non-fucosylated antibodies is expected to be a powerful and elegant approach to the design of the next generation therapeutic antibodies with improved efficacy. In this review, we discuss the importance of the oligosaccharides attached to the Fc region of therapeutic antibodies, especially regarding the inhibitory effect of fucosylated therapeutic antibodies on the efficacy of non-fucosylated counterparts in one medical agent. The impact of completely non-fucosylated therapeutic antibodies on therapeutic fields will be also discussed. 相似文献
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Dübel S 《Applied microbiology and biotechnology》2007,74(4):723-729
Recombinant antibody technology has revolutionized the development of antibody therapeutics. This minireview offers an overview
of enabling technologies and future prospects of this rapidly progressing field.
Remark: Trade names are copyright of distributing companies. 相似文献
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Andreas Loos Stefan Hillmer Josephine Grass Renate Kunert Jingyuan Cao David G. Robinson Ann Depicker Herta Steinkellner 《Plant biotechnology journal》2011,9(2):179-192
Seed‐specific expression is an appealing alternative technology for the production of recombinant proteins in transgenic plants. Whereas attractive yields of recombinant proteins have been achieved by this method, little attention has been paid to the intracellular deposition and the quality of such products. Here, we demonstrate a comparative study of two antiviral monoclonal antibodies (mAbs) (HA78 against Hepatitis A virus; 2G12 against HIV) expressed in seeds of Arabidopsis wild‐type (wt) plants and glycosylation mutants lacking plant specific N‐glycan residues. We demonstrate that 2G12 is produced with complex N‐glycans at great uniformity in the wt as well as in the glycosylation mutant, carrying a single dominant glycosylation species, GnGnXF and GnGn, respectively. HA78 in contrast, contains additionally to complex N‐glycans significant amounts of oligo‐mannosidic structures, which are typical for endoplasmic reticulum (ER)‐retained proteins. A detailed subcellular localization study demonstrated the deposition of both antibodies virtually exclusively in the extracellular space, illustrating their efficient secretion. In addition, although a KDEL‐tagged version of 2G12 exhibited an ER‐typical N‐glycosylation pattern, it was surprisingly detected in protein storage vacuoles. The different antibody variants showed different levels of degradation with hardly any degradation products detectable for HA78 carrying GnGnXF glycans. Finally, we demonstrate functional integrity of the HA78 and 2G12 glycoforms using viral inhibition assays. Our data therefore demonstrate the usability of transgenic seeds for the generation of mAbs with a controlled N‐glycosylation pattern, thus expanding the possibilities for the production of optimally glycosylated proteins with enhanced biological activities for the use as human therapeutics. 相似文献
8.
《MABS-AUSTIN》2013,5(3):413-415
Therapeutic monoclonal antibodies (mAbs) are currently being approved for marketing in Europe and the United States, as well as other countries, on a regular basis. As more mAbs become available to physicians and patients, keeping track of the number, types, production cell lines, antigenic targets, and dates and locations of approvals has become challenging. Data are presented here for 34 mAbs that were approved in either Europe or the United States (US) as of March 2012, and nimotuzumab, which is marketed outside Europe and the US. Of the 34 mAbs, 28 (abciximab, rituximab, basiliximab, palivizumab, infliximab, trastuzumab, alemtuzumab, adalimumab, tositumomab-I131, cetuximab, ibrituximab tiuxetan, omalizumab, bevacizumab, natalizumab, ranibizumab, panitumumab, eculizumab, certolizumab pegol, golimumab, canakinumab, catumaxomab, ustekinumab, tocilizumab, ofatumumab, denosumab, belimumab, ipilimumab, brentuximab) are currently marketed in Europe or the US. Data for six therapeutic mAbs (muromonab-CD3, nebacumab, edrecolomab, daclizumab, gemtuzumab ozogamicin, efalizumab) that were approved but have been withdrawn or discontinued from marketing in Europe or the US are also included. 相似文献
9.
Janice M. Reichert 《MABS-AUSTIN》2012,4(3):413-415
Therapeutic monoclonal antibodies (mAbs) are currently being approved for marketing in Europe and the United States, as well as other countries, on a regular basis. As more mAbs become available to physicians and patients, keeping track of the number, types, production cell lines, antigenic targets, and dates and locations of approvals has become challenging. Data are presented here for 34 mAbs that were approved in either Europe or the United States (US) as of March 2012, and nimotuzumab, which is marketed outside Europe and the US. Of the 34 mAbs, 28 (abciximab, rituximab, basiliximab, palivizumab, infliximab, trastuzumab, alemtuzumab, adalimumab, tositumomab-I131, cetuximab, ibrituximab tiuxetan, omalizumab, bevacizumab, natalizumab, ranibizumab, panitumumab, eculizumab, certolizumab pegol, golimumab, canakinumab, catumaxomab, ustekinumab, tocilizumab, ofatumumab, denosumab, belimumab, ipilimumab, brentuximab) are currently marketed in Europe or the US. Data for six therapeutic mAbs (muromonab-CD3, nebacumab, edrecolomab, daclizumab, gemtuzumab ozogamicin, efalizumab) that were approved but have been withdrawn or discontinued from marketing in Europe or the US are also included.Of the 28 mAbs currently marketed in the European Union or the US, 26 are marketed in Europe and 27 are marketed in the US, with 25 marketed in both regions (1 Of the 28 mAbs that are marketed in one or the other region, 43% (12/28) are produced in Chinese hamster ovary (CHO) cells, 25% (7/28) are produced in SP2/0 cells,2 18% (5/28) are produced in NS0 cells,3 and 7% (2/28) are produced in hybridomas. The remaining two products (ranibizumab, certolizumab pegol) are antigen-binding fragments (Fab) that are produced in E. coli. Humanized and human mAbs comprise 36% (10/28) and 32% (9/28) of the total, respectively, while 21% (6/28) are chimeric and 11% (3/28) are murine. Most (75%; 21/28) are canonical full-length mAbs. Of the 7 non-canonical mAbs, three (abciximab, ranibizumab, certolizumab pegol) are Fab, with one of these (certolizumab pegol) pegylated; two (tositumomab-I131, ibrituximab tiuxetan) are radiolabeled when administered to patients; one (brentuximab vedotin) is an antibody-drug conjugate (ADC); and one is bispecific (catumaxomab). Although 16 marketed mAbs target unique antigens, CD20 and tumor necrosis factor are each targeted by 4 mAbs, and epidermal growth factor receptor (EGFR) and vascular endothelial growth factor are each targeted by 2 mAbs. If approved, pertuzumab, which is undergoing regulatory review in Europe and the US as a treatment for breast cancer, would be one of 2 mAbs that target human epidermal growth factor receptor 2 on the market.Table 1. Therapeutic monoclonal antibodies marketed or in review in the European Union or United States
Open in a separate window*As of March 10, 2012. #Country-specific approval; approved under concertation procedure **Product manufactured for Phase 1 study in humans. Abbreviations: BLyS, B lymphocyte stimulator; C5, complement 5; CD, cluster of differentiation; CHO, Chinese hamster ovary; CTLA-4, cytotoxic T lymphocyte antigen 4; EGFR, epidermal growth factor receptor; EpCAM, epithelial cell adhesion molecule; Fab, antigen-binding fragment; GP glycoprotein; IL, interleukin; NA, not approved; PA, protective antigen; RANK-L, receptor activator of NFκb ligand; RSV, respiratory syncytial virus; TNF, tumor necrosis factor; VEGF, vascular endothelial growth factor. Sources: European Medicines Agency public assessment reports, United States Food and Drug Administration (drugs@fda), the international ImMunoGeneTics information system® (www.imgt.org/mAb-DB/index).In addition to the 28 mAbs currently marketed, six mAbs were approved in at least one country of Europe or in the US, but were subsequently withdrawn or discontinued from marketing for various reasons (4,5 Nebacumab (Centoxin®), a human IgM, was approved in The Netherlands, Britain, Germany and France during 1991 as a treatment for Gram-negative sepsis,6 but the product was subsequently withdrawn for safety, efficacy and commercial reasons.7 The murine anti-epithelial cell adhesion molecule (EpCAM) edrecolomab (Panorex®) was approved in Germany in 1995 as an adjuvant treatment for colon cancer, but subsequently withdrawn because of the product’s lack of efficacy.8 Daclizumab was first approved in 1997 for prophylaxis of acute organ rejection in patients receiving renal transplants, but the product was voluntarily withdrawn from the market in Europe effective January 1, 20099 and discontinued for the US market because of the availability of alternative therapy and the diminished market demand.10 The first ADC to be approved, gemtuzumab ozogamicin was marketed in the US for a decade before being voluntarily withdrawn in 2010. The product was approved under the accelerated approval mechanism as a treatment for acute myeloid leukemia (AML), but was withdrawn when a confirmatory clinical trial and post-approval use did not show evidence of clinical benefit in AML patients.11 Efalizumab (Raptiva®) was approved in the US and Europe in 2003 and 2004, respectively, as a treatment for adults with moderate to severe plaque psoriasis, but the product was voluntarily withdrawn from both markets in 2009 because of the risk of side effects, including progressive multifocal leukoencephalopathy.12,13Table 2. Therapeutic monoclonal antibodies withdrawn or discontinued from marketing in the European Union or United States
Open in a separate windowNote: Information current as of March 10, 2012. *European country-specific approval. Abbreviations: CD, cluster of differentiation; CHO, Chinese hamster ovary; EpCAM, epithelial cell adhesion molecule; IL, interleukin; NA, not approved. Sources: European Medicines Agency public assessment reports, United States Food and Drug Administration (drugs@fda), the international ImMunoGeneTics information system® (www.imgt.org/mAb-DB/index).The European Union and the US are not necessarily the first or only markets for therapeutic mAbs (14 Mogamulizumab is a defucosylated humanized anti-CC chemokine receptor 4 (CCR4) antibody developed by Kyowa Hakko Kirin Co Ltd.15 The mAb is undergoing regulatory review in Japan as a treatment for adult T-cell leukemia-lymphoma and peripheral T-cell lymphoma.Table 3. Therapeutic monoclonal antibodies marketed or in review outside the European Union or United States
Open in a separate windowNote: Information current as of March 10, 2012. Abbreviations: CCR, chemokine receptor; EGFR, epidermal growth factor receptor.The 35 marketed mAbs, most of which are canonical full-length IgG1, paved the way for the next generation of antibody-based therapeutics such as ADCs, bispecific antibodies, engineered antibodies, and antibody fragments or domains. The commercial pipeline includes ~350 mAbs now being evaluated in clinical studies around the world as treatments for many indications, including cancer, immunological disorders and infectious diseases.16 The compendium of marketed therapeutic antibodies may thus be substantially larger in the future. 相似文献
International non-proprietary name (Trade name) | Manufacturing cell line | Type | Target | First EU (US) approval year |
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Abciximab (Reopro®) | Sp2/0 | Chimeric IgG1κ Fab | GPIIb/IIIa | 1995# (1994) |
Rituximab (MabThera®, Rituxan®) | CHO | Chimeric IgG1κ | CD20 | 1998 (1997) |
Basiliximab (Simulect®) | Sp2/0 | Chimeric IgG1κ | IL2R | 1998 (1998) |
Palivizumab (Synagis®) | NS0 | Humanized IgG1κ | RSV | 1999 (1998) |
Infliximab (Remicade®) | Sp2/0 | Chimeric IgG1κ | TNF | 1999 (1998) |
Trastuzumab (Herceptin®) | CHO | Humanized IgG1κ | HER2 | 2000 (1998) |
Alemtuzumab (MabCampath, Campath-1H®) | CHO | Humanized IgG1κ | CD52 | 2001 (2001) |
Adalimumab (Humira®) | CHO | Human IgG1κ | TNF | 2003 (2002) |
Tositumomab-I131 (Bexxar®) | Hybridoma | Murine IgG2aλ | CD20 | NA (2003) |
Cetuximab (Erbitux®) | Sp2/0 | Chimeric IgG1κ | EGFR | 2004 (2004) |
Ibritumomab tiuxetan (Zevalin®) | CHO | Murine IgG1κ | CD20 | 2004 (2002) |
Omalizumab (Xolair®) | CHO | Humanized IgG1κ | IgE | 2005 (2003) |
Bevacizumab (Avastin®) | CHO | Humanized IgG1κ | VEGF | 2005 (2004) |
Natalizumab (Tysabri®) | NS0 | Humanized IgG4κ | α4-integrin | 2006 (2004) |
Ranibizumab (Lucentis®) | E. coli | Humanized IgG1κ Fab | VEGF | 2007 (2006) |
Panitumumab (Vectibix®) | CHO | Human IgG2κ | EGFR | 2007 (2006) |
Eculizumab (Soliris®) | NS0 | Humanized IgG2/4κ | C5 | 2007 (2007) |
Certolizumab pegol (Cimzia®) | E. coli | Humanized IgG1κ Fab, pegylated | TNF | 2009 (2008) |
Golimumab (Simponi®) | Sp2/0 | Human IgG1κ | TNF | 2009 (2009) |
Canakinumab (Ilaris®) | Sp2/0 | Human IgG1κ | IL1b | 2009 (2009) |
Catumaxomab (Removab®) | Hybrid hybridoma | Rat IgG2b/mouse IgG2a bispecific | EpCAM/CD3 | 2009 (NA) |
Ustekinumab (Stelara®) | Sp2/0 | Human IgG1κ | IL12/23 | 2009 (2009) |
Tocilizumab (RoActemra, Actemra®) | CHO | Humanized IgG1κ | IL6R | 2009 (2010) |
Ofatumumab (Arzerra®) | NS0 | Human IgG1κ | CD20 | 2010 (2009) |
Denosumab (Prolia®) | CHO | Human IgG2λ | RANK-L | 2010 (2010) |
Belimumab (Benlysta®) | NS0 | Human IgG1κ | BLyS | 2011 (2011) |
Raxibacumab (Pending) | NS0** | Human IgG1κ | B. anthrasis PA | NA (In review) |
Ipilimumab (Yervoy®) | CHO | Human IgG1κ | CTLA-4 | 2011 (2011) |
Brentuximab vedotin (Adcentris®) | CHO | Chimeric IgG1κ; conjugated to monomethyl auristatin E | CD30 | In review (2011) |
Pertuzumab (Pending) | CHO | Humanized IgG1κ | HER2 | In review (in review) |
International proprietary name (Trade name) | Manufacturing cell line | Type | Target | First EU (US) approval year |
---|---|---|---|---|
Muromonab-CD3 (Orthoclone OKT3®) | Hybridoma | Murine IgG2a | CD3 | 1986* (1986) |
Nebacumab (Centoxin®) | Hybridoma | Human IgM | Endotoxin | 1991*(NA) |
Edrecolomab (Panorex®) | Hybridoma | Murine IgG2a | EpCAM | 1995*(NA) |
Daclizumab (Zenapax®) | NS0 | Humanized IgG1κ | IL2R | 1999 (1997) |
Gemtuzumab ozogamicin (Mylotarg®) | NS0 | Humanized IgG4κ | CD33 | NA (2000) |
Efalizumab (Raptiva®) | CHO | Humanized IgG1κ | CD11a | 2004 (2003) |
International proprietary name (Trade name) | Manufacturing cell line | Type | Target | First approval year |
---|---|---|---|---|
Nimotuzumab (TheraCIM®, BIOMAB-EGFR®) | NS0 | Humanized IgG1κ | EGFR | 1999 |
Mogamulizumab | [Not found] | Humanized IgG1κ | CCR4 | In review in Japan |
10.
Production of vaccines and therapeutic antibodies for veterinary applications in transgenic plants: an overview 总被引:6,自引:0,他引:6
During the past two decades, antibodies, antibody derivatives and vaccines have been developed for therapeutic and diagnostic
applications in human and veterinary medicine. Numerous species of dicot and monocot plants have been genetically modified
to produce antibodies or vaccines, and a number of diverse transformation methods and strategies to enhance the accumulation
of the pharmaceutical proteins are now available. Veterinary applications are the specific focus of this article, in particular
for pathogenic viruses, bacteria and eukaryotic parasites. We focus on the advantages and remaining challenges of plant-based
therapeutic proteins for veterinary applications with emphasis on expression platforms, technologies and economic considerations. 相似文献
11.
Soomin Yoon Yong-Sung Kim Hyunbo Shim Junho Chung 《Biotechnology and Bioprocess Engineering》2010,15(5):709-715
Since the first monoclonal antibody, muromonab-CD3, was approved for therapeutic use in 1986, numerous molecules have been targeted using therapeutic antibody technology, resulting in 26 therapeutic antibodies being approved by the US FDA as of November, 2009. Initial concerns regarding antibody drugs focused on immunogenicity, short serum half-life, and weak efficacy. As the types of antibodies progressed from murine to chimeric, humanized, and fully human antibodies, great progress has been made in immunogenicity and in vivo instability issues. For example, humanized antibodies, such as bevacizumab, exhibit less than 0.2% immunogenicity and a 20 day serum half-life, which is comparable to native immunoglobulin. Some recently developed antibodies are exceedingly efficacious and have become first-line therapy for their target diseases. Here, we address and analyze all clinically approved therapeutic antibodies to date by discussing immunogenicity, half-life, and efficacy. 相似文献
12.
Gross M 《Current biology : CB》2001,11(14):R541-R542
Since the development of monoclonal antibodies twenty-five years ago, researchers and biotech companies have been looking to develop therapeutic uses for them. Michael Gross looks at some of the latest efforts. 相似文献
13.
With the advent of antibody fragments and alternative binding scaffolds, that are devoid of Fc-regions, strategies to increase the half-life of small proteins are becoming increasingly important. Currently, the established method is chemical PEGylation, but more elaborate approaches are being described such as polysialylation, amino acid polymers and albumin-binding derivatives. This article reviews the main strategies for pharmacokinetic enhancement, primarily chemical conjugates and recombinant fusions that increase apparent molecular weight or hydrodynamic radius or interact with serum albumin which itself has a long plasma half-life. We highlight the key chemical linkage methods that preserve antibody function and retain stability and look forward to the next generation of technologies which promise to make better quality pharmaceuticals with lower side effects. Although restricted to antibodies, all of the approaches covered can be applied to other biotherapeutics. 相似文献
14.
Elimination of the immunogenicity of therapeutic antibodies 总被引:4,自引:0,他引:4
Gilliland LK Walsh LA Frewin MR Wise MP Tone M Hale G Kioussis D Waldmann H 《Journal of immunology (Baltimore, Md. : 1950)》1999,162(6):3663-3671
The immunogenicity of therapeutic Abs limits their long-term use. The processes of complementarity-determining region grafting, resurfacing, and hyperchimerization diminish mAb immunogenicity by reducing the number of foreign residues. However, this does not prevent anti-idiotypic and anti-allotypic responses following repeated administration of cell-binding Abs. Classical studies have demonstrated that monomeric human IgG is profoundly tolerogenic in a number of species. If cell-binding Abs could be converted into monomeric non-cell-binding tolerogens, then it should be possible to pretolerize patients to the therapeutic cell-binding form. We demonstrate that non-cell-binding minimal mutants of the anti-CD52 Ab CAMPATH-1H lose immunogenicity and can tolerize to the "wild-type" Ab in CD52-expressing transgenic mice. This finding could have utility in the long-term administration of therapeutic proteins to humans. 相似文献
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16.
There are currently ~25 recombinant full-length IgGs (rIgGs) in the market that have been approved by regulatory agencies as biotherapeutics to treat various human diseases. Most of these are based on IgG1k framework and are either chimeric, humanized or human antibodies manufactured using either Chinese hamster ovary (CHO) cells or mouse myeloma cells as the expression system. Because CHO and mouse myeloma cells are mammalian cells, rIgGs produced in these cell lines are typically N-glycosylated at the conserved asparagine (Asn) residues in the CH2 domain of the Fc, which is also the case with serum IgGs. The Fc glycans present in these rIgGs are for the most part complex biantennary oligosaccharides with heterogeneity associated with the presence or the absence of several different terminal sugars. The major Fc glycans of rIgGs contain 0 or 1 or 2 (G0, G1 and G2, respectively) terminal galactose residues as non-reducing termini and their relative proportions may vary depending on the cell culture conditions in which they were expressed. Since glycosylation is strongly associated with antibody effector functions and terminal galactosylation may affect some of those functions, a panel of commercially available therapeutic rIgGs expressed in CHO cells and mouse myeloma cells were examined for their galactosylation patterns. The results suggest that the rIgGs expressed in CHO cells are generally less galactosylated compared to the rIgGs expressed in mouse myeloma cells. Accordingly, rIgGs produced in CHO cells tend to contain higher levels of G0 glycans compared with rIgGs produced in mouse myeloma cell lines. Despite the apparent wide variability in galactose content, adverse events or safety issues have not been associated with specific galactosylation patterns of therapeutic antibodies. Nevertheless, galactosylation may have an effect on the mechanisms of action of some therapeutic antibodies (e.g., effector pathways) and hence further studies to assess effects on product efficacy may be warranted for such antibodies. For antibodies that do not require effector functions for biological activity, however, setting a narrow specification range for galactose content may be unnecessary. 相似文献
17.
《MABS-AUSTIN》2013,5(3):385-391
There are currently ~25 recombinant full-length IgGs (rIgGs) in the market that have been approved by regulatory agencies as biotherapeutics to treat various human diseases. Most of these are based on IgG1k framework and are either chimeric, humanized or human antibodies manufactured using either Chinese hamster ovary (CHO) cells or mouse myeloma cells as the expression system. Because CHO and mouse myeloma cells are mammalian cells, rIgGs produced in these cell lines are typically N-glycosylated at the conserved asparagine (Asn) residues in the CH2 domain of the Fc, which is also the case with serum IgGs. The Fc glycans present in these rIgGs are for the most part complex biantennary oligosaccharides with heterogeneity associated with the presence or the absence of several different terminal sugars. The major Fc glycans of rIgGs contain 0 or 1 or 2 (G0, G1 and G2, respectively) terminal galactose residues as non-reducing termini and their relative proportions may vary depending on the cell culture conditions in which they were expressed. Since glycosylation is strongly associated with antibody effector functions and terminal galactosylation may affect some of those functions, a panel of commercially available therapeutic rIgGs expressed in CHO cells and mouse myeloma cells were examined for their galactosylation patterns. The results suggest that the rIgGs expressed in CHO cells are generally less galactosylated compared to the rIgGs expressed in mouse myeloma cells. Accordingly, rIgGs produced in CHO cells tend to contain higher levels of G0 glycans compared with rIgGs produced in mouse myeloma cell lines. Despite the apparent wide variability in galactose content, adverse events or safety issues have not been associated with specific galactosylation patterns of therapeutic antibodies. Nevertheless, galactosylation may have an effect on the mechanisms of action of some therapeutic antibodies (e.g., effector pathways) and hence further studies to assess effects on product efficacy may be warranted for such antibodies. For antibodies that do not require effector functions for biological activity, however, setting a narrow specification range for galactose content may be unnecessary. 相似文献
18.
The binding sites on human IgG1 for human Fc gamma receptor (Fc gamma R) I, Fc gamma RIIa, Fc gamma RIIb, Fc gamma RIIIa and neonatal FcR have been mapped. A common set of IgG1 residues is involved in binding to all Fc gamma Rs, while Fc gamma RII and Fc gamma RIII utilize distinct sites outside this common set. In addition to residues which abrogated binding to the Fc gamma R, several positions were found which improved binding only to specific Fc gamma Rs or simultaneously improved binding to one type of Fc gamma R and reduced binding to another type. Selected IgG1 variants with improved binding to Fc gamma RIIIa were then tested in an in vitro antibody-dependent cellular cytotoxicity (ADCC) assay and showed an enhancement in ADCC when either peripheral blood mononuclear cells or natural killer cells were used. 相似文献
19.
The formation of covalently linked, high molecular weight protein aggregates has been thought to play an important role in opacification of the human lens. Antisera were used in Western blot analysis to demonstrate the involvement of all major classes of lens proteins (alpha, beta and gamma crystallin; the major intrinsic membrane polypeptide) in covalent aggregation. Of these classes, aggregation of gamma and beta crystallins via intermolecular disulfide bonding and aggregation of the major intrinsic membrane polypeptide via intermolecular, non-disulfide bonding were more pronounced in cataractous as compared with normal lenses. 相似文献
20.
Controlled glycosylation of therapeutic antibodies in plants 总被引:5,自引:0,他引:5
Tekoah Y Ko K Koprowski H Harvey DJ Wormald MR Dwek RA Rudd PM 《Archives of biochemistry and biophysics》2004,426(2):266-278
Recombinant therapeutic monoclonal antibodies (mAb) can be expressed, assembled, and glycosylated in plants. Transgenic plants, producing anti-rabies mAb and anti-colorectal cancer mAb, were obtained from Agrobacterium-mediated transformation. The heavy chain (HC) of anti-rabies mAb was fused to the Lys-Asp-Glu-Leu (KDEL) endoplasmic reticulum retention signal whereas the HC of anti-colorectal cancer mAb was not fused to the KDEL sequence. Gel release of glycans and detection by high-performance liquid chromatography (HPLC), together with computer assisted analysis and matrix-assisted laser desorption/ionization time-of-flight (MALD-TOF) mass spectrometry, revealed that the plant-derived anti-rabies mAb with KDEL contained mainly oligomannose type N-glycans while the plant-derived anti-colorectal cancer mAb carried mainly biantennary glycans with and without a pentose sugar, that is thought to be xylose. This finding indicates that the KDEL sequence can affect the N-glycosylation processing of antibody in plant cells. The plant-derived mAbs with addition of a KDEL sequence did not contain any of the known antigenic glycan epitopes that are frequently found in other plant glycans or in mammalian-derived mAbs. The altered glycosylation on both plant-derived mAbs did not affect the activities that are required for therapy. These results indicate that plant genetic engineering could provide an effective and inexpensive means to control the glycosylation of therapeutic proteins such as mAbs, by the addition of a KDEL signal as a regulatory element. 相似文献