Carbohydrate Analysis of a Chimeric Recombinant Monoclonal Antibody by Capillary Electrophoresis with Laser-Induced Fluorescence Detection

Ma, Stacey; Nashabeh, Wassim

doi:10.1021/ac990376z

Cited by 131 publications

(114 citation statements)

References 39 publications

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“…The glycan structure of rituximab was determined by using capillary electrophoresis (CE) with laser-induced fluorescence detection after digestion with various glycosidases (14). The released glycans were analyzed after derivatization with 8-aminopyrene-1,3,6-trisulfonic acid, trisodium salt (APTS) on a Beckman Coulter P͞ACE MDQ CE system equipped with an argon-ion laser with an excitation wavelength of 488 nm and an emission band-pass filter of 520 Ϯ 10 nm.…”

Section: Methodsmentioning

confidence: 99%

Crystalline monoclonal antibodies for subcutaneous delivery

Yang

Shenoy

Disttler

et al. 2003

Proc. Natl. Acad. Sci. U.S.A.

142

130

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Therapeutic applications for mAbs have increased dramatically in recent years, but the large quantities required for clinical efficacy have limited the options that might be used for administration and thus have placed certain limitations on the use of these agents. We present an approach that allows for s.c. delivery of a small volume of a highly concentrated form of mAbs. Batch crystallization of three Ab-based therapeutics, rituximab, trastuzumab, and infliximab, provided products in high yield, with no detectable alteration to these proteins and with full retention of their biological activity in vitro. Administration s.c. of a crystalline preparation resulted in a remarkably long pharmacokinetic serum profile and a dose-dependent inhibition of tumor growth in nude mice bearing BT-474 xenografts (human breast cancer cells) in vivo. Overall, this approach of generating high-concentration, low-viscosity crystalline preparations of therapeutic Abs should lead to improved ease of administration and patient compliance, thus providing new opportunities for the biotechnology industry.

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Section: Methodsmentioning

confidence: 99%

Crystalline monoclonal antibodies for subcutaneous delivery

Yang

Shenoy

Disttler

et al. 2003

Proc. Natl. Acad. Sci. U.S.A.

142

130

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“…The use 460 of CE-LIF as a method of glycan separation of monoclonal antibodies has been thoroughly discussed previously and the separation of glycans by this method for the elucidation of structural isomers has proven effective. 12,31,37 However, as these CE-LIF studies in the literature have been compared to separation of glycans by HI-LIC-UPLC, the separation of structural isomers will not be discussed further here. Besides the CE system used in this study, other commercially available general purpose CE units can also be used in a similar manner.…”

Section: Q2mentioning

confidence: 99%

“…Rapid analysis methods are beneficial for high throughput; however, results may vary depending on the separation technique selected. Current techniques for profiling, characterization and analysis of glycans include hydrophilic 70 liquid chromatography (HILIC), 4-7 reverse phase chromatography (RP), 8,9 capillary electrophoresis (CE), [10][11][12] mass spectrometry (MS), [13][14][15][16][17] nuclear magnetic resonance (NMR) 18,19 amongst others.…”

Section: Introductionmentioning

confidence: 99%

Comparison of separation techniques for the elucidation of IgG N-glycans pooled from healthy mammalian species

Adamczyk

Tharmalingam-Jaikaran

Schomberg

et al. 2014

Carbohydrate Research

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a b s t r a c tThe IgG N-glycome provides sufficient complexity and information content to serve as an excellent source for biomarker discovery in mammalian health. Since oligosaccharides play a significant role in many biological processes it is very important to understand their structure. The glycosylation is cell type specific as well as highly variable depending on the species producing the IgG. We evaluated the variation of N-linked glycosylation of human, bovine, ovine, equine, canine and feline IgG using three orthogonal glycan separation techniques: hydrophilic interaction liquid chromatography (HILIC)-UPLC, reversed phase (RP)-UPLC and capillary electrophoresis with laser induced fluorescence detection (CE-LIF). The separation of the glycans by these high resolution methods yielded different profiles due to diverse chemistries. However, the % abundance of structures obtained by CE-LIF and HILIC-UPLC were similar, whereas the analysis by RP-UPLC was difficult to compare as the structures were separated by classes of glycans (highly mannosylated, fucosylated, bisected, fucosylated and bisected) resulting in the co-elution of many structures. The IgGs from various species were selected due to the complexity and variation in their N-glycan composition thereby highlighting the complementarity of these separation techniques.

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“…Radio labeling of the released glycans, and analyzing them by a combination of paper electrophoresis and gel-filtration chromatography [13], with normal phase high-performance liquid chromatography (HPLC) [14,15] or anion-exchange HPLC [16] coupled with a fluorescence detector have also been described. Additionally, capillary electrophoresis coupled with different detectors has been described [17][18][19]. More recently, several methods using mass spectrometry (MS) have been developed.…”

mentioning

confidence: 99%

Electrospray ionization quadrupole ion-mobility time-of-flight mass spectrometry as a tool to distinguish the lot-to-lot heterogeneity in N-glycosylation profile of the therapeutic monoclonal antibody trastuzumab

Damen

Chen

Chakraborty

et al. 2009

J. Am. Soc. Mass Spectrom.

126

102

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Monoclonal antibodies are typically glycosylated at asparagine residues in the Fc domain, and glycosylation heterogeneity at the Fc sites is well known. This paper presents a method for rapid analysis of glycosylation profile of the therapeutic monoclonal antibody trastuzumab from different production batches using electrospray quadrupole ion-mobility time-of-flight mass spectrometry (ESI-Q-IM-TOF). The global glycosylation profile for each production batch was obtained by a fast LC-MS analysis, and comparisons of the glycoprofiles of trastuzumab from different lots were made based on the deconvoluted intact mass spectra. Furthermore, the heterogeneity at each glycosylation site was characterized at the reduced antibody level and at the isolated glycopeptide level. The glycosylation site and glycan structures were confirmed by performing a time-aligned-parallel fragmentation approach using the unique dual-collision cell design of the instrument and the incorporated ion-mobility separation function. Four different production batches of trastuzumab were analyzed and compared in terms of global glycosylation profiles as well as the heterogeneity at each glycosylation site. The results show that each batch of trastuzumab shares the same types of glycoforms but relative abundance of each glycoforms is varied. (J Am Soc Mass

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Carbohydrate Analysis of a Chimeric Recombinant Monoclonal Antibody by Capillary Electrophoresis with Laser-Induced Fluorescence Detection

Cited by 131 publications

References 39 publications

Crystalline monoclonal antibodies for subcutaneous delivery

Crystalline monoclonal antibodies for subcutaneous delivery

Comparison of separation techniques for the elucidation of IgG N-glycans pooled from healthy mammalian species

Electrospray ionization quadrupole ion-mobility time-of-flight mass spectrometry as a tool to distinguish the lot-to-lot heterogeneity in N-glycosylation profile of the therapeutic monoclonal antibody trastuzumab

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