Separation window dependent multiple injection (SWDMI) for large scale analysis of therapeutic antibody N-glycans

Zsuzsanna Kovács, Máté Szarka, Márton Szigeti, A. Guttman

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

There is a growing demand in the biopharmaceutical industry for large scale N-glycosylation analysis of biotherapeutics, especially monoclonal antibodies. To fulfill this high throughput analysis requirement with single column separation systems in most instances require finishing the entire analysis cycle including conditioning, injection and separation between sample injections. While in liquid chromatography it represents a challenge, multiple sample injection in capillary electrophoresis has already been demonstrated for one or two sample components by utilizing the concept of introducing sequential sample and buffer zones into the capillary tubing before the start of the separation process. It was also demonstrated in CE-MS mode, mostly to follow one sample component, identified by precise mass measurement. Here we introduce a novel multiple injection approach for rapid large scale capillary electrophoresis analysis of samples with biopharmaceutical interest supporting multicomponent optical detection with laser induced fluorescence. In Separation Window Dependent Multiple Injection (SWDMI) mode, the samples are consecutively injected in predefined time intervals, based on the window that covers the separation of all sample components. As a practical example, this newly developed SWDMI protocol was applied to rapid and large scale analysis of APTS labeled monoclonal antibody N-glycans using a short (20 cm effective length) capillary column. Full analysis of 96 samples (injected from a well plate) was obtained in 4 h, in contrast to consecutive individual separation cycle processing of the same samples that required 12 h.

Original languageEnglish
Pages (from-to)367-370
Number of pages4
JournalJournal of Pharmaceutical and Biomedical Analysis
Volume128
DOIs
Publication statusPublished - Sep 5 2016

Fingerprint

Polysaccharides
Injections
Antibodies
Capillary Electrophoresis
Capillary electrophoresis
Capillary Tubing
Therapeutics
Monoclonal Antibodies
Glycosylation
Liquid Chromatography
Liquid chromatography
Tubing
Industry
Buffers
Lasers
Fluorescence
Throughput
Processing

Keywords

  • Biopharmaceuticals
  • High throughput analysis
  • N-glycans

ASJC Scopus subject areas

  • Analytical Chemistry
  • Drug Discovery
  • Pharmaceutical Science
  • Spectroscopy
  • Clinical Biochemistry

Cite this

Separation window dependent multiple injection (SWDMI) for large scale analysis of therapeutic antibody N-glycans. / Kovács, Zsuzsanna; Szarka, Máté; Szigeti, Márton; Guttman, A.

In: Journal of Pharmaceutical and Biomedical Analysis, Vol. 128, 05.09.2016, p. 367-370.

Research output: Contribution to journalArticle

@article{4d44035816de4a70a66167a66282f3b0,
title = "Separation window dependent multiple injection (SWDMI) for large scale analysis of therapeutic antibody N-glycans",
abstract = "There is a growing demand in the biopharmaceutical industry for large scale N-glycosylation analysis of biotherapeutics, especially monoclonal antibodies. To fulfill this high throughput analysis requirement with single column separation systems in most instances require finishing the entire analysis cycle including conditioning, injection and separation between sample injections. While in liquid chromatography it represents a challenge, multiple sample injection in capillary electrophoresis has already been demonstrated for one or two sample components by utilizing the concept of introducing sequential sample and buffer zones into the capillary tubing before the start of the separation process. It was also demonstrated in CE-MS mode, mostly to follow one sample component, identified by precise mass measurement. Here we introduce a novel multiple injection approach for rapid large scale capillary electrophoresis analysis of samples with biopharmaceutical interest supporting multicomponent optical detection with laser induced fluorescence. In Separation Window Dependent Multiple Injection (SWDMI) mode, the samples are consecutively injected in predefined time intervals, based on the window that covers the separation of all sample components. As a practical example, this newly developed SWDMI protocol was applied to rapid and large scale analysis of APTS labeled monoclonal antibody N-glycans using a short (20 cm effective length) capillary column. Full analysis of 96 samples (injected from a well plate) was obtained in 4 h, in contrast to consecutive individual separation cycle processing of the same samples that required 12 h.",
keywords = "Biopharmaceuticals, High throughput analysis, N-glycans",
author = "Zsuzsanna Kov{\'a}cs and M{\'a}t{\'e} Szarka and M{\'a}rton Szigeti and A. Guttman",
year = "2016",
month = "9",
day = "5",
doi = "10.1016/j.jpba.2016.06.002",
language = "English",
volume = "128",
pages = "367--370",
journal = "Journal of Pharmaceutical and Biomedical Analysis",
issn = "0731-7085",
publisher = "Elsevier",

}

TY - JOUR

T1 - Separation window dependent multiple injection (SWDMI) for large scale analysis of therapeutic antibody N-glycans

AU - Kovács, Zsuzsanna

AU - Szarka, Máté

AU - Szigeti, Márton

AU - Guttman, A.

PY - 2016/9/5

Y1 - 2016/9/5

N2 - There is a growing demand in the biopharmaceutical industry for large scale N-glycosylation analysis of biotherapeutics, especially monoclonal antibodies. To fulfill this high throughput analysis requirement with single column separation systems in most instances require finishing the entire analysis cycle including conditioning, injection and separation between sample injections. While in liquid chromatography it represents a challenge, multiple sample injection in capillary electrophoresis has already been demonstrated for one or two sample components by utilizing the concept of introducing sequential sample and buffer zones into the capillary tubing before the start of the separation process. It was also demonstrated in CE-MS mode, mostly to follow one sample component, identified by precise mass measurement. Here we introduce a novel multiple injection approach for rapid large scale capillary electrophoresis analysis of samples with biopharmaceutical interest supporting multicomponent optical detection with laser induced fluorescence. In Separation Window Dependent Multiple Injection (SWDMI) mode, the samples are consecutively injected in predefined time intervals, based on the window that covers the separation of all sample components. As a practical example, this newly developed SWDMI protocol was applied to rapid and large scale analysis of APTS labeled monoclonal antibody N-glycans using a short (20 cm effective length) capillary column. Full analysis of 96 samples (injected from a well plate) was obtained in 4 h, in contrast to consecutive individual separation cycle processing of the same samples that required 12 h.

AB - There is a growing demand in the biopharmaceutical industry for large scale N-glycosylation analysis of biotherapeutics, especially monoclonal antibodies. To fulfill this high throughput analysis requirement with single column separation systems in most instances require finishing the entire analysis cycle including conditioning, injection and separation between sample injections. While in liquid chromatography it represents a challenge, multiple sample injection in capillary electrophoresis has already been demonstrated for one or two sample components by utilizing the concept of introducing sequential sample and buffer zones into the capillary tubing before the start of the separation process. It was also demonstrated in CE-MS mode, mostly to follow one sample component, identified by precise mass measurement. Here we introduce a novel multiple injection approach for rapid large scale capillary electrophoresis analysis of samples with biopharmaceutical interest supporting multicomponent optical detection with laser induced fluorescence. In Separation Window Dependent Multiple Injection (SWDMI) mode, the samples are consecutively injected in predefined time intervals, based on the window that covers the separation of all sample components. As a practical example, this newly developed SWDMI protocol was applied to rapid and large scale analysis of APTS labeled monoclonal antibody N-glycans using a short (20 cm effective length) capillary column. Full analysis of 96 samples (injected from a well plate) was obtained in 4 h, in contrast to consecutive individual separation cycle processing of the same samples that required 12 h.

KW - Biopharmaceuticals

KW - High throughput analysis

KW - N-glycans

UR - http://www.scopus.com/inward/record.url?scp=84975507098&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84975507098&partnerID=8YFLogxK

U2 - 10.1016/j.jpba.2016.06.002

DO - 10.1016/j.jpba.2016.06.002

M3 - Article

C2 - 27337190

AN - SCOPUS:84975507098

VL - 128

SP - 367

EP - 370

JO - Journal of Pharmaceutical and Biomedical Analysis

JF - Journal of Pharmaceutical and Biomedical Analysis

SN - 0731-7085

ER -