Structural Characterization of the N-Glycans of a Humanized Anti-CD18 Murine Immunoglobulin G

Ip, Charlotte C. Yu; Miller, William J.; Silberklang, Melvin; Mark, George E.; Ellis, Ronald W.; Huang, Lingyi; Glushka, John; Vanhalbeek, H.; Zhu, Jing; Alhadeff, Jack A.

doi:10.1006/abbi.1994.1055

Cited by 38 publications

(9 citation statements)

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“…The initial mammalian endoplasmic reticulum pathway, including trimming of the terminal glucose and mannose residues, seems operative. The trimming process appears to follow a linear pathway with the exception of two different forms of the Man 7 GlcNAc 2 (M7.2 and M7.1), also observed for native insect glycoproteins (7) and IgG 4 from NS/0 cells (41). The presence of two M7 forms suggests the possibility of an alternative processing pathway (7) such as one that yields the M7.2 oligosaccharide through the action of endo-␣-mannosi- …”

Section: Comparison Of Hplc Oligosaccharide Profiles In the Presence mentioning

confidence: 71%

“…A comparison of the carbohydrate structures from IgG 2a secreted from insect cells with those of the recombinant IgG 4 secreted from NS/0 mouse myeloma cells is also informative (41). While the proportion of complex structures obtained from NS/0 was higher (46 versus 35%), a number of the complex and hybrid structures obtained from the two sources were similar.…”

Section: Comparison Of Hplc Oligosaccharide Profiles In the Presence mentioning

confidence: 99%

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Differential N-Glycan Patterns of Secreted and Intracellular IgG Produced in Trichoplusia ni Cells

Hsu

Takahashi

Tsukamoto

et al. 1997

Journal of Biological Chemistry

113

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Insect cells have been widely utilized as hosts for the production of numerous glycoproteins through the baculovirus expression system (1, 2). Native insect cell glycoproteins also serve as models for developmental processes in eucaryotes (3, 4). While the structure, synthesis, and function of oligosaccharides in mammalian glycoproteins are well characterized, information on the carbohydrate structures and processing pathways present in insect cells is limited and sometimes contradictory (5-7). The oligosaccharides in glycoproteins can play critical roles in cellular targeting, structural stability, resistance to proteolysis, immunogenicity, and circulatory halflife (8, 9). With insects representing more than half of the animal species classified (5), there is a need to obtain more information on the carbohydrate structures and processing of glycoproteins from insect cells.Many initial studies of N-glycans in insect cell-derived heterologous glycoproteins indicated the presence of only high mannose-type or short truncated, paucimannosidic oligosaccharides, 1 sometimes containing L-fucosyl 2 residues (6, 7). These observations confirmed the earlier studies of endogenous insect cell glycoproteins which were similarly found to lack complex carbohydrate structures (10 -12). It was presumed that insect cells did not possess the capacity to synthesize complex-type oligosaccharides since the levels of sialyl-, galactosyl-, and N-acetylglucosamine transferases were found to be insignificant (12).In contrast, studies with the recombinant human plasminogen indicated that insect cells could synthesize complex Nlinked oligosaccharides (13). Several more recent studies on homologous glycoproteins also have indicated that certain insect cell lines can synthesize hybrid and complex oligosaccharides. Honeybee venom phospholipase A 2 (PLA 2 ) 3 was found to

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Section: Comparison Of Hplc Oligosaccharide Profiles In the Presence mentioning

confidence: 71%

Section: Comparison Of Hplc Oligosaccharide Profiles In the Presence mentioning

confidence: 99%

Differential N-Glycan Patterns of Secreted and Intracellular IgG Produced in Trichoplusia ni Cells

Hsu

Takahashi

Tsukamoto

et al. 1997

Journal of Biological Chemistry

113

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“…48,63,[113][114][115][116][117][118][119][120][121][122][123][124][125][126][127] Oligosaccharides of high mannose have also been reported for several antibodies. 33,62,97,103,111,[128][129][130][131] Sialic acid is normally present at a very low level for Fc oligosaccharides. 47,101,102,111,[132][133][134][135][136] While different subtypes of IgGs 137,138 and cell culture conditions 33,93,130,134,[139][140][141][142][143] can affect oligosaccharide structures, cell line 45,46,137,138,141,144,145 ...…”

Section: Chemical Modifications Disulfide Bondsmentioning

confidence: 99%

Heterogeneity of Monoclonal Antibodies

Liu

Gaza-Bulseco

Faldu

et al. 2008

Journal of Pharmaceutical Sciences

424

343

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“…The structure of the N-glycans corresponding to the numbered peaks (1 to 10) in Figure 5 have all been identified in earlier work. 35 The predominant species were biantennary, complex N-glycans that terminate in either N-acetyl glucosamine (GlcNAc) on both arms (peak 4), or a single arm with galactose (Gal) on the other arm (peak 6). No significant levels (< 10%) of N-glycans terminating with sialic acid were detected.…”

Section: Gi Ycosylationmentioning

confidence: 99%

Characterization of a recombinant antibody produced in the course of a high yield fed‐batch process

Robinson

Chan

et al. 1994

Biotech & Bioengineering

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Many mammalian cell fed-batch processes rely on maintaining the cells in a viable and productive state for extended periods of time in order to reach high final concentrations of secreted protein. In the work described herein, a nonamplified NSO cell line was transfected with a vector expressing a recombinant human anti-HIV gp 120 monoclonal antibody (Mab) and a selectable marker, glutamine synthetase. A fed-batch process was developed which improved product yields tenfold over the yields reached in batch culture. In this case, the clone was cultured for a period of 22 days and produced 0.85 g Mab/L. To gauge the effect of extended culture lifetime on product quality, biochemical characteristics of MAb isolated from different time points in the fed-batch culture were determined. The apparent molecular weight of the MAb was constant throughout the course of the culture. Isoelectric focusing revealed four major charged species, with a fifth more acidic species appearing later in the culture. The antigen binding kinetics were constant for MAb isolated throughout the culture period. Glycosylation analysis, on the other hand, revealed that MAb produced later in the culture contained greater percentages of truncated N-acetylglucosamine and highmannose N-glycans. Possible contributions to this underglycosylated material from either cell lysis or synthesis from noviable cells were found to be negligible. Instead, the viable cells appeared to be secreting more truncated and high mannose MAb glycoforms as the culture progressed.

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Structural Characterization of the N-Glycans of a Humanized Anti-CD18 Murine Immunoglobulin G

Cited by 38 publications

References 0 publications

Differential N-Glycan Patterns of Secreted and Intracellular IgG Produced in Trichoplusia ni Cells

Differential N-Glycan Patterns of Secreted and Intracellular IgG Produced in Trichoplusia ni Cells

Heterogeneity of Monoclonal Antibodies

Characterization of a recombinant antibody produced in the course of a high yield fed‐batch process

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