Immunogenicity of N-Glycolylneuraminic Acid-Containing Carbohydrate Chains of Recombinant Human Erythropoietin Expressed in Chinese Hamster Ovary Cells

Noguchi, Akira; Mukuria, Chege J.; Suzuki, Emiko; Naiki, Masaharu

doi:10.1093/oxfordjournals.jbchem.a124721

Cited by 97 publications

(68 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This exceeds the low proportion (1% to 5%) of NeuGc previously reported for recombinant proteins produced by CHO cells (Hokke et al, 1995), and may be specific to this cell line. As CMP-NeuAc hydroxylase is not present in humans (Irie and Suzuki, 1998;Muchmore et al, 1989), NeuGc is considered to be potentially immunogenic (Noguchi et al, 1995). Therefore, N-glycan substitution with NeuGc is undesirable, even for recombinant IgG proteins where sialylation is reduced by steric hindrance.…”

Section: Comparative Analysis Of N-glycosylation In Gs-ns0 and Gs-chomentioning

confidence: 99%

Metabolic control of recombinant protein N‐glycan processing in NS0 and CHO cells

Baker

Rendall

Hills

et al. 2001

Biotech & Bioengineering

167

116

View full text Add to dashboard Cite

Chinese hamster ovary and murine myeloma NS0 cells are currently favored host cell types for the production of therapeutic recombinant proteins. In this study, we compared N-glycan processing in GS-NS0 and GS-CHO cells producing the same model recombinant glycoprotein, tissue inhibitor of metalloproteinases 1. By manipulation of intracellular nucleotide-sugar content, we examined the feasibility of implementing metabolic control strategies aimed at reducing the occurrence of murine-specific glycan motifs on NS0-derived recombinant proteins, such as Galalpha1,3Galbeta1,4GlcNAc. Although both CHO and NS0-derived oligosaccharides were predominantly of the standard complex type with variable sialylation, 30% of N-glycan antennae associated with NS0-derived TIMP-1 terminated in alpha1,3-linked galactose residues. Furthermore, NS0 cells conferred a greater proportion of terminal N-glycolylneuraminic (sialic) acid residues as compared with the N-acetylneuraminic acid variant. Inclusion of the nucleotide-sugar precursors, glucosamine (10 mM, plus 2 mM uridine) and N-acetylmannosamine (20 mM), in culture media were shown to significantly increase the intracellular pools of UDP-N-acetylhexosamine and CMP-sialic acid, respectively, in both NS0 and CHO cells. The elevated UDP-N-acetylhexosamine content induced by the glucosamine/uridine treatment was associated with an increase in the antennarity of N-glycans associated with TIMP-1 produced in CHO cells but not N-glycans associated with TIMP-1 from NS0 cells. In addition, elevated UDP-N-acetylhexosamine content was associated with a slight decrease in sialylation in both cell lines. The elevated CMP-sialic acid content induced by N-acetylmannosamine had no effect on the overall level of sialylation of TIMP-1 produced by both CHO and NS0 cells, although the ratio of N-glycolylneuraminic acid:N-acetylneuraminic acid associated with NS0-derived TIMP-1 changed from 1:1 to 1:2. These data suggest that manipulation of nucleotide-sugar metabolism can promote changes in N-glycan processing that are either conserved between NS0 and CHO cells or specific to either NS0 cells or CHO cells.

show abstract

Section: Comparative Analysis Of N-glycosylation In Gs-ns0 and Gs-chomentioning

confidence: 99%

Metabolic control of recombinant protein N‐glycan processing in NS0 and CHO cells

Baker

Rendall

Hills

et al. 2001

Biotech & Bioengineering

167

116

View full text Add to dashboard Cite

show abstract

“…A HD antibody, which was obtained from a patient with lung cancer, recognized the Neu5Gc epitope of ESPO and EPOGIN [5]. Therefore, it was con firmed that the EIA using GM3(Neu5Gc) as the antigen can detect antibodies to Neu5Gc epitope of ESPO and EPOGIN.…”

Section: Resultsmentioning

confidence: 99%

“…Thus, even 1 % Neu5Gc as in rHuE PO may possibly stimulate antibody production and lead to an allergic-like serum sickness in patients. We therefore first immunized chickens with rHuEPO as a model sys tem [5], Two chickens responded weakly to the Neu5Gc epitope of rHuEPO but another 2 did not respond signifi cantly, while those immunized with fetuin, which con tains 6% Neu5Gc, produced significantly high titers of HD antibodies. This immunization was carried out by one subcutaneous administration of the antigen with Freund's complete adjuvant.…”

Section: Discussionmentioning

confidence: 99%

“…Recently, however, an anaphylac tic reaction to rHuEPO look place in one Spanish patient during intravenous administration and where rHuEPOspecific IgE antibody was detected [4], One of the antigenic determinants in rHuEPO may be Neu5Gc. Indeed, the immunization of chickens (who like humans do not synthesize Neu5Gc) with rHuEPO led to a production of low titer antibody against Neu5Gc epitope [5], Human cancerous tissues and some chicken lympho mas express Neu5Gc in their sialoglycoconjugates, and this expression frequently stimulates the production of the so-called Hanganutziu-Deicher (HD) antibody which recognizes Neu5Gc moiety of the sialoglycoconjugates [6,7].…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Failure of Human Immunoresponse to N-Glycolylneuraminic Acid Epitope Contained in Recombinant Human Erythropoietin

Noguchi

Mukuria

Suzuki

et al. 1996

Nephron

Self Cite

View full text Add to dashboard Cite

Recombinant human erythropoietin (rHuEPO) was produced by Chinese hamster ovary cells and commercially distributed to hospitals by two pharmaceutical companies in Japan (‘ESPO’ by Kirin Brewery Co. Ltd., and Sankyo Co. Ltd., and ‘EPOGIN’ by Chugai Pharmaceutical Co. Ltd.) These products contained about 1% N-glycolylneuraminic acid (Neu5Gc) in total sialic acid content. Since humans do not synthesize Neu5Gc, successive injection of Neu5Gc-containing products was feared to lead to allergic-like symptoms. Therefore, serum levels of antibodies to Neu5Gc epitope in 90 patients who received repeated i.v. injections of ESPO or EPOGIN were determined by an enzyme immunoassay using Neu5Gcα2-3Galβ1 -4Glc-Cer, GM3(Neu5Gc), as an antigen and compared with those in 100 healthy persons. Either no or low antibody levels were detected in both groups with no significant difference. In 40 patients who received s.c. injections of ESPO or EPOGIN, serum HD antibody levels were determined before and after weekly therapeutic injections carried out for one to several months, but no significant elevations were detected in all patients. The above results indicated that therapeutic administration of rHuEPO to patients with chronic renal failure is safe from allergic-like side effects associated with the production of Neu5Gc-specific antibodies, and it was concluded that Neu5Gc epitope of rHuEPO is minimally antigenic in humans.

show abstract

“…In an antibody-based product, the N-linked oligosaccharide is sequestered in the CH2 domain and therefore not considered to possess immunogenic potential. However, the O-glycosylation near the hinge region of CNTO736 is relatively more exposed and is considered to be potentially more immunogenic (Noguchi et al, 1995). This will especially be true if non-human type sialic acid, N-glycolyl-Neuraminic acid (Neu5Gc), synthesized by mouse myeloma cells were present in the O-glycosylated species.…”

Section: Product Quality Is Dependent On the Host Cell In Which It Ismentioning

confidence: 99%

Development of mammalian production cell lines expressing CNTO736, a glucagon like peptide‐1‐MIMETIBODY^TM: Factors that influence productivity and product quality

Dorai

Nemeth

Cammaart

et al. 2009

Biotech & Bioengineering

View full text Add to dashboard Cite

In an attempt to develop a high producing mammalian cell line expressing CNTO736, a Glucagon like peptide-1-antibody fusion protein (also known as a Glucagon like peptide-1 MIMETIBODY), we have noted that the N-terminal GLP-1 portion of the MIMETIBODY was susceptible to proteolytic degradation during cell culture, which resulted in an inactive product. Therefore, a number of parameters that had an effect on productivity as well as product quality were examined. Results suggest that the choice of the host cell line had a significant effect on the overall product quality. Product expressed in mouse myeloma host cell lines had a lesser degree of proteolytic degradation and variability in O-linked glycosylation as compared to that expressed in CHO host cell lines. The choice of a specific CHOK1SV derived clone also had an effect on the product quality. In general, molecules that exhibited minimal N-terminal clipping had increased level of O-linked glycosylation in the linker region, giving credence to the hypothesis that O-linked glycosylation acts to protect against proteolytic degradation. Moreover, products with reduced potential for N-terminal clipping had longer in vivo serum half-life. These findings suggest that early monitoring of product quality should be an essential part of production cell line development and therefore, has been incorporated in our process of cell line development for this class of molecules.

show abstract

Immunogenicity of N-Glycolylneuraminic Acid-Containing Carbohydrate Chains of Recombinant Human Erythropoietin Expressed in Chinese Hamster Ovary Cells

Cited by 97 publications

References 0 publications

Metabolic control of recombinant protein N‐glycan processing in NS0 and CHO cells

Metabolic control of recombinant protein N‐glycan processing in NS0 and CHO cells

Failure of Human Immunoresponse to N-Glycolylneuraminic Acid Epitope Contained in Recombinant Human Erythropoietin

Development of mammalian production cell lines expressing CNTO736, a glucagon like peptide‐1‐MIMETIBODY^TM: Factors that influence productivity and product quality

Contact Info

Product

Resources

About

Immunogenicity of N-Glycolylneuraminic Acid-Containing Carbohydrate Chains of Recombinant Human Erythropoietin Expressed in Chinese Hamster Ovary Cells

Cited by 97 publications

References 0 publications

Metabolic control of recombinant protein N‐glycan processing in NS0 and CHO cells

Metabolic control of recombinant protein N‐glycan processing in NS0 and CHO cells

Failure of Human Immunoresponse to N-Glycolylneuraminic Acid Epitope Contained in Recombinant Human Erythropoietin

Development of mammalian production cell lines expressing CNTO736, a glucagon like peptide‐1‐MIMETIBODYTM: Factors that influence productivity and product quality

Contact Info

Product

Resources

About

Development of mammalian production cell lines expressing CNTO736, a glucagon like peptide‐1‐MIMETIBODY^TM: Factors that influence productivity and product quality