Articles by Maria Ekemohn

FabRICATOR®-HPLC for Automated Antibody Subunit Analysis

December 17, 2019 | References |

Middle-level analysis is a universally accepted analytical strategy for rapid characterization of antibodies. The FabRICATOR® (IdeS)enzyme specifically digests IgG below the hinge, generating  fragments that are suitable for high-resolution mass spectrometry. By immobilizing the FabRICATOR enzyme in an HPLC column format, a fully automated on-column digestion of IgG is possible.
 

In a recent article written by the Genovis team and published in Chromatography Today, an LC-MS workflow for automated middle-level analysis of monoclonal antibodies (mAbs) and mAbs-based biologics is described and evaluated. FabRICATOR-HPLC was shown to facilitate easy on-line sample preparation for middle-level LC-MS. On-column digestion of mAbs followed by on-column reduction and analysis by RP-HPLC and MS yielded results indistinguishable from those produced using an off-line sample preparation protocol.
 

“With minimal carry-over, low sample requirements and a tolerance for a wide concentration range, this method is very versatile. It is well suited for both routine analysis as well as more advanced applications such as automatic monitoring of mAb CQAs during production in a bioreactor.”
 

The full article is available here:

Nägeli et al., 2020. On-column digestion of mAbs for automated middle-level analysis by LC-MS. Chromatograpy Today
 

Read more about FabRICATOR-HPLC and automated antibody subunit analysis

SmartEnzymes™ Simplify Characterization of Charge Variants by Native Mass Spectrometry

November 19, 2019 | References |

Monoclonal antibodies (mAbs) are complex macromolecules that undergo a wide range of post-translational modifications that can result in charge heterogeneity. Ion exchange (IEX) chromatography is considered the gold standard for characterization of charge variants. This approach separates charge variants and allows for the collection of isoforms for protein identification by mass spectrometry (MS). However, the collection of multiple fractions often needs to be combined with top-down and bottom-up characterization methods, which is both resource and time consuming.

 

In a recent study by LeBlanc et al., (2019) from LFB Biotechnologies, a newly developed cation exchange column technology was evaluated for its ability to separate charge variants under native conditions with direct coupling to MS. In total, three different mAbs with high basicisoelectric points (pI) and a monoclonal antibody reference material from NIST were analyzed in both their native and proteolyzed forms. The fragments were obtained using the IgG-specific proteases FabRICATOR®, which digests below the hinge, and FabALACTICA®, which digests above the hinge.

 

The column was shown to separate mAbs with high basicpI, demonstrating that mAbs having a strong retention on cation exchange media could be separated. A good repeatability was achieved in terms of resolution, recovery and retention times. Overall, the results demonstrate how SmartEnzymes have made mAb charge variant characterization, which was formerly challenging, more user-friendly with an easy-to-replicate protocol.

 

Read more about FabRICATOR and FabALACTICA.

 

Leblanc, Y. et al., 2019. A generic method for intact and subunit level characterization of mAb charge variants by native mass spectrometry, Journal of Chromatography B. doi: https://doi.org/10.1016/j.jchromb.2019.121814

Monitoring Glycation Levels on Bispecific Biologics using FabRICATOR®

November 18, 2019 | References |

Bispecific monoclonal antibodies (BsAbs) are multi-functioning and complex biologics with the ability to recognize two different epitopes for improved therapeutic properties. Characterizing protein modifications such as glycation on biologics is vital to ensure consistency in stability and function. The structural complexity of BsAbs requires robust analytical methods, where conventional top-down and bottom-up strategies may lack in sensitivity or even introduce further modifications. Middle-level analysis using site-specific proteases such as GingisKHAN®(Kgp) and FabRICATOR®(IdeS) is an intermediate strategy that enables complementary analysis of intact or reduced Fab and Fc fragments.

 

In a recent article by Gstöttner et al. (2019) from Leiden University together with Roche Pharma Technical Development, the authors analyzed a BsAb for protein modification levels, N- and C-terminal sequencing and modification localization using top-down, middle-level and bottom-up strategies. The BsAb was analyzed for changes in glycation levels over time using middle-up FT-ICR MS on Fc/2, LC and Fd’ fragments obtained by FabRICATOR digestion. The scientists also localized glycation hot spots on the heavy chain backbone of the FabRICATOR-digested BsAb using sequential in source decay (ISD) MALDI fragmentation.

 

Using FabRICATOR in a novel middle-up MS strategy, the scientists were able to analyze all antibody subunits in a single high-resolution mass spectrum. By implementing the method in a forced-glycation experiment, changes in glycation levels were successfully monitored over time. The authors were also able to localize several glycation hot spots by intact top-down and FabRICATOR-assisted middle-down analyses. The use of middle-level strategies in combination with conventional MS-based methods successfully provided complementary data for monitoring the level of glycation.

 

Learn more about FabRICATOR and our other proteases.

 

Gstöttner, C. et al., 2019. Monitoring glycation levels of a bispecific monoclonal antibody at subunit level by ultrahigh resolution MALDI FT-ICR mass spectrometry. mAbs. doi: 10.1080/19420862.2019.1682403.

SmartEnzymes™ in a new approach to characterize ADCs

October 25, 2019 | Applications, References |

Antibody drug conjugates (ADCs) consist of monoclonal antibodies chemically linked to a cytotoxic agent. The target specificity of the monoclonal antibody in combination with the potency of the cytotoxic drug make ADCs promising therapeutic agents. However, the molecules are often complex, making evaluation of the quality attributes for the ADC challenging.

 

In order to characterize the ADCs, the predominant analysis of choice is peptide mapping with reversed-phase liquid chromatography (RPLC) coupled to mass spectrometry. However, the sample preparation steps in a bottom-up approach are often time-consuming and a comprehensive view of ADCs with different sequence variants and post-translational modifications is lacking.

 

In this recently published article by Chen et al., a middle-down RPLC-MS strategy with electron transfer disscociation (ETD) was developed to analyze lysine and cysteine conjugated ADCs at the subunit level. FabRICATOR® (IdeS) and GingisKHAN® (KGB) were used to generate the subunits. FabRICATOR digests below the hinge, generating F(ab’)2 and Fc/2 fragments, and GingisKHAN digests above the hinge, generating intact Fab and Fc fragments. For the deglycosylation, the IgG-specific endoglycosidase GlycINATOR® (EndoS2) was used.

 

This middle-down approach enabled high-resolution evaluation of several ADC quality attributes at the subunit level, including drug to antibody ratio (DAR), conjugation sites and micro-variants. The approach shows great potential for investigating quality attributes during the development and characterization of novel ADCs.

 

Read more about FabRICATOR, GingisKHAN and GlycINATOR.

 

Chen, B et al., 2019. Middle-Down Multi-Attribute Analysis of Antibody-Drug Conjugates with Electron Transfer Dissociation. Anal. Chem. 91(18). 11661-11669.

Characterizing ADCs using FabRICATOR and middle-down MS

September 24, 2019 | Applications, Products |

The process of characterizing an antibody drug conjugate (ADC) requires the evaluation of critical quality attributes including primary sequence analysis and drug conjugation assessment. Addressing glycoprofile determination as well as drug load distribution and drug-to-antibody ratio (DAR) is however challenging using peptide mapping. In addition, further challenges arise from the increased hydrophobicity of the ADC and the risk of drug-linker dissociation in an MS/MS experiment.

   

In an article by Hernandez-Alba et al. (2019) the authors characterized a site-specific ADC using middle-down MS. They combined three different fragmentation strategies for improved sequence coverage and drug conjugation assessment. The site-specific ADC (DAR=4) was digested using FabRICATOR and reduced to generate homogenous Fc/2, Fd’ and LC fragments for analysis by multiple ion activation techniques. By combining MS/MS data obtained with HDC (hydrodynamic chromatography), ETD (electron-transfer dissociation) and UVPD (ultraviolet photodissociation) fragmentation modes, the scientists obtained valuable information with the advantages of minimal sample preparation and analysis time using middle-down MS. 

   

The UVPD mode showed better performance compared to ETD and HDC. This indicated that the performance of this activation technique was unaffected by the hydrophobicity of the ADC. The complementarity between UVPD and ETD was further highlighted for drug conjugation assessment by allowing primary sequence validation and accurate identification of drug conjugation and glycosylation sites. These results highlight the potential of middle-down MS as a complement in next-generation strategies for the characterization of  mAb-based compounds including ADCs. 

 
 

Read more about FabRICATOR and Applications of FabRICATOR.

   

Hernandez-Alba, O. et al., 2019. A Case Study to Identify the Drug Conjugation Site of a Site-Specific Antibody-Drug-Conjugate using Middle-Down Mass Spectrometry. American Society for Mass Spectrometry, 30(8). pp. 1-11. 

Introducing our new CFO, Johny Humaloja

September 9, 2019 | Genovis Team, Interview |

 

Johny joined the Genovis team in August, and he brings more than 25 years of financial experience to the company. We are very excited to have you in the team, Johny! Welcome!

 

Tell us a bit about yourself

I live in Malmö, in the southern part of Sweden and I have two daughters. I have a cat named Zelda. On my free time I like to play hockey with my friends and go out for a ride on my road bike.

 

If you were to describe yourself using only one word – what word would that be?

Committed

 

Tell us about your previous working life

I have more than 18 years of finance leadership experience in the life science industry, mainly from working in US public listed companies such as Biogen and Boston Scientific. Focus has been on the Nordic region, to develop businesses and execute operational improvements. At Biogen I got the opportunity to work as Plant Controller and run all the financial activities at their large-scale manufacturing plant in North Carolina.

 

What will your main focus be here at Genovis?

Day-to-day business and make sure to enable Genovis to continue to grow and expand the business model. It will be key for me to understand the different parts of the organization and give adequate financial information and support. Financial areas to focus on will be;internal control, project management, statutory accounting, taxes, manage outsourced services, and external reporting.

 

What do you believe will be the biggest opportunity in your new position as CFO?

I believe I can utilize skills and experiences in sales/commercial and production area to find new ways to do business and provide financial and strategic leadership.

 

4 quick questions:

Coffee or tea?

Coffee – Zoegas

Aerosmith or Depeche Mode?

Depeche Mode

Ice cream or candy?

Chocolate Ice cream

Soccer or ice hockey?

Hockey, Carolina Hurricanes

FragIT™ kit in Study Evaluating IgG Charge

September 2, 2019 | References |

 

Protein charge is a fundamental property that influences both the ability to interact with other molecules and the structure, solubility and stability of the protein.

In this collaborative work by Boehringer Ingelheim Pharmaceuticals, Janssen BioTherapeutics and University of New Hampshire, the charge measurements of twelve monoclonal IgG and their F(ab’)2 and Fc fragments are presented. Besides other interesting findings, it is suggested that mAb charge measurements is valuable when it comes to selecting candidate molecules for development.

FragIT kit was used to achieve the IgG subunits before charge evaluation. This kit consists of spin columns of an immobilized version of the FabRICATOR (IdeS) enzyme for antibody digestion and spin columns for affinity binding of the Fc fragments. Using this kit, the Fc and F(ab’)2 fragments were easily separated from each other, with no enzyme in the final preparation.

 

For more information about FragIT kit, please visit the following page:

FragIT kit

 

The full-text paper is available online:

Yang et al., 2019. IgG Charge: Practical and Biological Implications. Antibodies, 8(1), 24

 

The Digested Guide to ASMS 2019

Many of Genovis’ customers have submitted abstracts for ASMS 2019 in Atlanta, in which the SmartEnzymes have been used. Below is a digested guide to these abstracts so that you can plan your days at ASMS.

To read the full poster abstracts, visit the Online Planner for ASMS, and paste the abstract titel in the search field.

 

Sunday, June 2

New Enzymatic Workflows for Analysis of O-Glycosylated Biopharmaceuticals

Andreas Nägeli

Thermo Fisher Scientific User Meeting

Pharma/BioPharma Breakout Session 9:30 AM – 12.15 PM

Marquis Ballroom C

Thermo-user-meeting-blog

Monday, June 3

Improved middle-down characterization of antibodies using multiple ion activation techniques and Proton Transfer Reaction on a modified Orbitrap mass spectrometer 

Romain Huguet

MP 676, Poster Session 10.30 AM – 2.30 PM

FabRICATOR® and GingisKHAN®

 

Direct Determination of Antibody Chain Pairing by Top-Down Mass Spectrometry using Electron Capture Dissociation and Ultraviolet Photodissociation 

Jared Shaw

Room B302-305, Oral Presentation 3.50 PM – 4.10 PM

FabRICATOR® and GingisKHAN®

 

Unraveling a complex immunoprotein profile in multiple myeloma with middle-down de novo sequencing and native mass spectrometry 

Valerie J Winton

Room A411-412, Oral Presentation 9.30 AM – 9.50 AM

FabRICATOR®

 

Middle Down Approach for the Characterization of Monoclonal Antibodies After Ides Digestion and ETD Fragmentation

Colin Wynne

MP 782, Poster Session 10.30 AM – 2.30 PM

FabRICATOR®

 

Tuesday, June 4

Analysis of Zika Viral Polyprotein N- and O-glycosylation Using a Novel Lectin-chemoenzymatic Enrichment 

Shuang Yang

TP 655, Poster Session 10.30 AM – 2.30 PM

OpeRATOR® and GlycOCATCH™

 

Comprehensive Characterization of Antibody Drug Conjugates Enabled by Top-down and Middle-down Mass Spectrometry Strategies

Eli J Larson

TP 601, Poster Session 10.30 AM – 2.30 PM

GlycINATOR®FabRICATOR® and GingisKHAN®

 

Wednesday, June 5

Analysis of O-glycosylated Biopharmaceuticals using an O-glycan dependent Endoprotease and LC-MS

Andreas Nägeli

WP 334, Poster Session 10.30 AM – 2.30 PM

OpeRATOR® and OglyZOR®

 

MALDI-in-source decay FT-ICR MS for top-down and middle-down characterization of monoclonal antibodies 

Simone Nicolardi

WP 032, Poster Session 10.30 AM – 2.30 PM

FabRICATOR® and GingisKHAN®

 

Application of a label-free and domain-specific free thiol method in monoclonal antibody characterization

Yi Pu

WP 040, Poster Session 10.30 AM – 2.30 PM

FabALACTICA® and FabRICATOR®

 

Collision induced unfolding experiments to decipher the structural regions of a hybrid monoclonal antibody

Thomas Botzanowski

WP 481, Poster Session 10.30 AM – 2.30 PM

IgGZERO®

 

Thursday, June 6

Intact and Subunit Mass Analysis Using Native Ion Exchange Chromatography Coupled to an Orbitrap Mass Spectrometer 

Qian Liu

ThP 663, Poster Session 10.30 AM – 2.30 PM

FabRICATOR®

 

Ultra-Fast Analysis of Intact Proteins Using SPE- TOF 

Kevin McCann

ThP 149, Poster Session 10.30 AM – 2.30 PM

FabRICATOR®

 

Isomeric linkage determination of Sialic acid on O-glycopeptides using O-protease and LC-MS/MS 

Jieqiang Zhong

ThP 071, Poster Session 10.30 AM – 2.30 PM

OpeRATOR®

Free Thiols using FabRICATOR® and FabALACTICA®

In biopharmaceutical product development and manufacturing, free thiol content is one of the product quality attributes of interest as its presence could impact structure, stability and function of the product.

At Biogen, Yi Pu et al have optimized a label-free LC (UV) / MS method for free thiol quantification at a subunit level of IgG1 and IgG4. The new method, which is based on a method developed by Faid et al*, was compared to two conventional approaches, Ellman’s assay and peptide mapping.

It is very challenging to identify free thiol forms by mass spectrometry at the intact antibody level. By combining the highly specific proteolytic enzymes FabALACTICA (IgdE) and FabRICATOR (IdeS) the authors generated the subunits Fab, hinge and Fc/2, suited for confident mass determination. The subunits were subsequently separated on a polyphenyl reversed phase column in order to separate free thiol forms from their corresponding disulphide bond-linked form. A baseline or near baseline separation was obtained making it possible to calculate the free thiol content on each subunit.

The result of the quantification of free thiols from all three methods were comparable and showed similar trends even though the peptide mapping approach generally gave a higher free thiol content.

The authors conclude that compared to Ellman’s assay, the subunit approach is more sensitive, requires less sample and provides domain-specific information of the free thiol content. Compared to peptide mapping, the subunit method is faster, less labour intensive and lacks dependence on labelling efficiency. Finally, it demonstrated promise in the quantification of free thiols in a high throughput manner with domain specific information available.

The developed method has successfully been applied to several in-house IgG1 mAbs with different hydrophobicity and isoelectric points.

 

*V. Faid Y. Leblanc N. Bihoreau G. Chevreux Middle-up analysis of monoclonal antibodies after combined IgdE and IdeS hinge proteolysis: Investigation of free sulfhydryls, J. Pharm. Biomed. Anal. 149 (2018) 541-546, https://doi.org/10.1016/j.jpba.2017.11.046

 

For more information on FabRICATOR and FabALACTICA please visit the following pages:

The full text paper is available online:

The Digested Guide to ASMS 2018

 

Several researchers have submitted abstracts for ASMS 2018 in San Diego, in which Genovis’ SmartEnzymes have been used. Below is a selection of these abstracts.

To read the poster abstracts, visit the Online Planner for ASMS, and paste the abstract titel in the search field.

Monday June 4 

10:30AM-2:30PM: Poster Session

MP 045 – GlyCLICK

Development of NISTmAb-derived homogeneous antibody-drug conjugate (ADC) standards

Shanhua Lin1; Terry Zhang2; Brian Agnew3; Trina Mouchahoir4; John Schiel4
1Thermo Fisher Scientific, Sunnyvale, CA; 2Thermo Fisher Scientific, San Jose, CA; 3Thermo Fisher Scientific, Eugene, Oregon; 4NIST, Gaithersburg, MD

MP 046 – FabRICATOR

Discovery and confirmation of glucuronylation as a new acidic post-translational modification on therapeutic monoclonal antibodies

Yuetian Yan1; Anita Liu1; Shunhai Wang1; Thomas Daly1; Ning Li1
1Regeneron Pharmaceuticals, Tarrytown, NY

MP 049 – FabRICATOR

Ultrasensitive Characterization of Size and Charge Heterogeneity of Therapeutic Monoclonal Antibodies by Native Mass Spectrometry

Shunhai Wang1; Yuetian Yan1; Anita Liu1; Thomas Daly1; Ning Li1
1Regeneron Pharmaceuticals, Tarrytown, NY

MP 464 – FabALACTICA

Quantitative UPLC-MSE analysis of disulfide bonds and free sulfhydryls in monoclonal antibodies using IgdE protease assisted digestion

Jeroen de Keijzer1; Peter van Maurik1; Anja Boumeester1; Emile van Corven1; Gideon Oudgenoeg1
1Bioceros, Utrecht, Netherlands

 

Tuesday June 5 

10:30AM-2:30PM: Poster Session

TP 624 – FabRICATOR

Avoiding method induced heterogeneity in the analysis of heterogeneity of monoclonal antibodies using Mass Spectrometry after Single Site Proteolysis

Gideon Oudgenoeg1; Anja Boumeester2; Peter van Maurik2; Jeroen de Keijzer2; Emile van Corven2
1Bioceros, Utrecht, Netherlands; 2Bioceros, Utrecht, Netherlands

 

Wednesday June 6 

10:30AM-2:30PM: Poster Session

WP056 – FabRICATOR

Characterizing and Quantitating Therapeutic Antibody Multimer Degradation Using Affinity Capture Mass

Neha Srikumar1; Wenjing Li1; Robert Tchelepi1; Chen Gu1; Diego Ellerman1; Greg A Lazar1; Yichin Liu1John C. Tran1
1Genentech Inc., South San Francisco, CA

WP 327  – GingisKHAN

Comprehensive Domain-Specific [Fc vs. Fab] N-glycosylation Analysis of Therapeutic Proteins

Charles Nwosu1; Shuangqi Sally Liu2; Lei Wang2; May Zhu2; Anne Kowal2
1Takeda Pharmaceuticals International Co, Cambridge, MA; 2Takeda Pharmaceuticals International Co., Cambridge, MA

WP 340 – FabULOUS

Analysis of Fragmented Porcine Immunoglobulin G (IgG) by MALDI-MS and UPLC-ESI-MS

HELENE PERREAULT1; Claudia Nelson2
1University of manitoba, Winnipeg; 2University of Manitoba, Winnipeg, MB

WP 342 – OpeRATOR and GlycOCATCH

A Novel O-glycoprotease with applications in O-glycan Analysis using mass spectrometry

Rolf Lood1, 2; Maria Nordgren1; Fredrik Leo1; Stephan Björk1; Malin Mejáre1Fredrik Olsson1
1Genovis AB, Lund, Sweden; 2Department of Infectious Diseases, Lund University, Lund, Sweden

WP 350 – OpeRATOR

Deciphering complex o-glycosylation: solid-phase chemoenzymatic cleavage and enrichment

Shuang Yang1; Philip Onigman2; Jonathan Sjogren2; Wells W. Wu3; Rong-fong Shen3; John Cipollo1
1LBP, CBER, FDA, Silver Spring, MD; 2Genovis AB Inc., Boston, MA; 3FBR, CBER, FDA, Silver Spring, MD

WP 676 –FabRICATOR, GlycINATOR, IgGZERO

Monitoring Critical Quality Attributes: Core Fucosylation of N-glycans using an Integrated Subunit LC/MS Workflow Method

Nilini S Ranbaduge1; Henry Y Shion1; Ying Qing Yu1; Weibin Chen1
1Waters Corporation, Milford, MA

WP 678 – FabRICATOR

In-depth Characterization of the Heterogeneous Dimerization Interfaces of A Monoclonal Antibody: from Subdomain Level to Residue Level

Yuetian Yan1; Shunhai Wang1Thomas Daly1; Ning Li1
1Regeneron Pharmaceuticals, Tarrytown, NY

W 691 – OpeRATOR, OglyZOR and SialEXO

LC-MS Characterization of Complex Glycoproteins

Amber Peariso1; Jason X. Tang1
1Eli Lilly & Company, Indianapolis, IN

WP 692 – FabRICATOR

Rapid Identity Assays for mAb Development, Production Control and Release

Anja Resemann1; Waltraud Evers1Yue Ju2; Guillaume Tremintin2; Detlev Suckau1
1Bruker Daltonics, Bremen, Germany; 2Bruker Daltonics, Billerica, MA

 

Thursday June 7 

10:10AM-10:30PM: Oral Presentation

Oral Presentation – FabRICATOR

Classification of Plasma Cell Disorders by 21 Tesla FT-ICR Top-Down and Middle-Down MS/MS Analysis of Monoclonal Immunoglobulins in Human Serum

Lidong He1, 2; Lissa C Anderson2; David R Barnidge3; David L Murray4; Surendra Dasari4; Angela Dispenzieri4; Christopher L Hendrickson1, 2; Alan G Marshall1, 2
1Florida State University, Tallahassee, FL; 2National High Magnetic Field Laboratory, Tallahassee, FL; 3The Binding Site, Rochester, MN; 4Mayo Clinic, Rochester, MN

 

10:30AM-2:30PM: Poster Session

ThP 003 – FabRICATOR

LC-MS in Combination with Multiple Enzymatic Digestion for Sequence Variant Identification in Support of Cell Line Development

Renpeng Liu1; Lintao Wang1; Alexandru C. Lazar1
1ImmunoGen, Waltham, MA

ThP 017 – GingisKHAN

Consortium for Top-Down Proteomics Inter-laboratory Study for Characterizing Monoclonal Antibodies (mAbs) by Top-Down Mass Spectrometry

Kristina Srzentic1; Luca Fornelli1; Yury Tsybin2; Joseph Loo3; Jeffrey Agar4; Julia Chamot-Rooke5Paul Danis6; Ying Ge7; David Goodlett8; Neil Kelleher1; Ljiljana Pasa Tolic9; Lloyd Smith7; Timothy Toby1; Konstantin Nagornov2; JENNIFER BRODBELT10; Sylvester Greer10; Mathieu Dupré5; David Clarke11; Ziqing Lin7; Kim Haselmann12; Christopher Hendrickson13; Lidong He13; Donald Hunt14; Jared Shaw9; Wendy Sandoval15; Richa Sarin16; Detlev Suckau17; Yuri E.M. van der Burgt18; Norelle Wildburger19; Nicolas L. Young20; Alain Beck21; John Yates22; Jolene Diedric22; Sneha Chatterjee23; Frank Sobott24; Anton Kozhinov2; ALAN G. MARSHALL13; LISSA C. ANDERSON13; Natalia Gasilova25; Laure Menin25; Neil Quebbenamm3; Sung Hwan Yoon26; Josh Hinkle14; Simone Nicolardi18; Matthew V. Holt20; Yunqiu Chen16; Nicholas Schmitt4
1Northwestern University, Evanston, IL; 2Spectroswiss Sàrl, Lausanne, Switzerland; 3UCLA, Los Angeles, CA; 4Northeastern University, Boston, MA; 5Institute Pasteur, Paris, France; 6Eastwoods Consulting, Boylston, MA; 7University of Wisconsin, Madison, WI; 8University of Maryland, Baltimore, Baltimore; 9PNNL, Richland, WA; 10University of Texas at Austin, Austin, TX; 11Edinburgh University, Edinburgh, United Kingdom; 12Novo Nordisk, Malov, Denmark; 13National High Magnetic Field Laboratory, Tallahassee, FL; 14University of Virginia, Charlottesville, VA; 15Genentech, Inc., South San Francisco, CA; 16Biogen Inc, Cambridge, MA; 17Bruker Daltonik GmbH, Bremen, Germany; 18Leiden University Medical Centre, Leiden, Netherlands; 19Washington University, St. Louis, St. Louis, MO; 20Baylor College of Medicine, Houston, TX; 21Centre d’immunologie Pierre Fabre, Saint-Julien-en-Genevois, France; 22The Scripps Research Institute, La Jolla, CA; 23University of Antwerp, Antwerp, Belgium; 24University of Leeds, Leeds, United Kingdom; 25Ecole Polytechnique Fédérale de Lausanne, Ch-1015 Lausanne, Switzerland; 26University of Maryland, Baltimore, MD

ThP 028 – FabRICATOR

Top- and Middle-Down CE-ESI-MS Analysis of Intact mAbs Using the ZipChip Coupled to a Fusion Lumos ETD Mass Spectrometer

Tricia C. Ho1; Erik J. Soderblom1; Erin Redman2; Greg M. Waitt1; M. Arthur Moseley1
1Duke University School of Medicine, Proteomics and Metabolomics Shared Resource, Durham, NC; 2908 Devices, Inc., Carrboro, NC

ThP 419 – FabRICATOR

Quantitation of Misincorporations: Strategies and System Suitability

Kathleen Cornelius1; Olga Friese1; Mary Denton2; Jason Rouse2
1Pfizer, Inc, Chesterfield, MO; 2Pfizer Inc., Andover, MA

 

2:30 -2:50 PM: Oral Presentation

Ballroom 20D – FabRICATOR, GingisKHAN, GlycINATOR and IgGZERO

A Suite of Liquid Chromatography Strategies Coupled Online to Top-down High-resolution Mass Spectrometry for Comprehensive Analysis of Antibody Drug Conjugates

Bifan Chen1; Ziqing Lin1; Qingge Xu1; Cexiong Fu2; Qunying Zhang2; Ying Ge1
1University of Wisconsin-Madison, Madison, Wisconsin; 2Abbvie Inc., North Chicago, IL