Research & Development for Biopharmaceuticals



Ludger blood spot




Ludger's biopharmaceutical R&D is aimed at understanding the structure-function of biopharmaceuticals in terms of glycosylation. The goal of this programme is to improve drug development by the following steps:

  • Changing glycosylation of drugs to increase safety and efficacy
  • Implementing Quality by Design glycoprofiling workflows to maximise efficiency of drug monitoring (using LongBow)





Collaborative Projects:

Ludger-H2020-Marie Curie

2017-2021

A4B - Analytics for Biologics - EU Horizon 2020 research and innovation programme (Marie Sklodowska-Curie grant agreement No 765502): a research training network on qualitative and quantitative analysis and purification of therapeutic proteins

This pan European consortium focusses on the qualitative and quantitative analysis and purification of therapeutic proteins. This 4-year project begins in October 2017 and a PhD student (ESR) will be based at Ludger for three years. Our PhD student will focus on the development of automatable and high throughput techniques to determine site specificity and quantification of N and O glycan profiles of EPO and TNF-AB. The project will build upon a recently developed product, VTAG, used to analyze and relatively quantifyglycan types on the Fc receptor of monoclonal antibodies without releasing the glycan. Glycans from the therapeutic proteins will be analysed using UHPLC based hydrophilic interaction chromatography, and a glycopeptide fluorophore label will be optimized for MS and CGE-LIF analysis by chemical modification. Additionally, quantitative glycopeptide standards will be prepared and finally the assay will be automated and validated according to ICHQ2(R1) guidelines.

www.analytics4biologics.eu




Ludger-Glycoenzymes

2015-2020

Glycoenzymes for Bioindustries

Glycoenzymes are responsible for the biosynthesis of all glycans and glycoconjugates and as such provide a rich source of biocatalysts for industrial applications; however, few glycoenzymes are commercially available. The Glycoenzymes for Bioindustries project has been created to generate a panel of glycoenzymes which can be commercialised for glycoscience and Industrial Biotechnology applications. In this 60 month project, approximately 1000 different enzymes will be produced along with a comprehensive database of benchmarked performance data. Selected examples will be produced at industrial specification for evaluation by industrial partners. The goal of the project is to enable the increased use of glycoenzymes, shaping future R&D and transforming industrial processes.

The project has been awarded funding by Innovate UK, BBSRC and EPSRC. The consortium is comprised of the following partners: University of Manchester, John Innes Centre, University of Newcastle, Institute of Food Research, Ludger Ltd (industrial partner), Biocatalysis Ltd (industrial partner), and Prozomix Ltd (sub contractor).

www.glycoenzymes.com




Ludger-King's College-IBCarb

Jan 2017-Sept 2017

GlycoShape: Defining and designing altered IgE glycoforms

Academic supervisors: Dr Gerd Wagner & Dr Sophia Karagiannis (King's College London). Industrial partner: Ludger Ltd.

Immunoglobulin E (IgE)-based antibodies are currently under investigation as novel immunotherapeutics for the treatment of cancer [1]. IgE is a heavily glycosylated glycoprotein, whose glycans make up more than 10% of its molecular mass [2]. Manipulating the glycan structure of IgE therefore offers a unique opportunity to alter the biological properties of IgE-based antibodies. However, while it is clear that the number and nature of IgE glycans significantly affect the biological and pharmacological activity of IgE-based antibodies, it is not clear, how.

In the GlycoShape project, we seek to clarify the role of glycans for IgE activity, and to exploit this knowledge for the development of altered IgE glycoforms. Bringing together expertise in IgE pharmacology (Karagiannis), chemical inhibitors of glycan biosynthesis (Wagner) and high-performance glycoanalytics (Ludger Ltd), we will provide proof-of-concept for a cost effective, flexible, operationally simple and potentially generic method for the glycoengineering of recombinant therapeutic glycoproteins.

REFERENCES
[1] Josephs DH, Spicer JF, Karagiannis P, Gould HJ, Karagiannis SN. IgE immunotherapy: a novel concept with promise for the treatment of cancer. MAbs 2014, 6, 54.
[2] Plomp R, Hensbergen PJ, [...] Wuhrer M. Site-Specific N-Glycosylation Analysis of Human Immunoglobulin E. J. Proteome Res. 2014, 13, 536.






Ludger-High Glycan

2011-2016

HIGH GLYCAN - EU FP7-funded programme for development of high throughput glycomics technology.

Dysfunctional glycosylation is associated with a wide range of diseases including cancer, diabetes, as well as congenital, cardiovascular, immunological and infectious disorders. Furthermore, it is widely accepted that glycosylation of biopharmaceuticals can impact safety and therapeutic efficacy. Despite the strong evidence for measuring glycosylation in disease and biologicals, glycomics is significantly lagging behind genomics and proteomics, mainly due to the absence of high-throughput analytical methods which can reliably quantify a multitude of glycan structures in complex biological samples. The HighGlycan programme has been set up to address this by developing high thoughput glycan analysis using HPLC, MS and CGE-LIF technologies.

www.highglycan.eu




Ludger-University of Sheffield

2015-2019
UK BBSRC iCASE award PhD student - Sheffield University


bbsrc









Ludger-University of Reading

2013-2017
UK BBSRC iCASE award PhD student - Reading University

bbsrc












Relevant posters and papers:


Ludger WCBP2018 FSH characterisation poster

Biopharmaceutical follicle-stimulating hormone (FSH) characterisation: monitoring Glycosylation Critical Quality Attributes (GCQAs) using a procainamide labelling system for structural glycan analysis and LC-ESI-QTOF for glycopeptide mapping

Thomson RI, Gardner RA, Kozak RP, Smith J, Strohfeldt K, Osborn HMI, Spencer DI
Presented at: WCBP 2018: 22nd Symposium on the Interface of Regulatory and Analytical Sciences for Biotechnology Health Products
Washington D.C., United States. January 30-February 1st 2018





Ludger WCBP2018 Sialylation poster

Sialylation: A Critical Glycosylation Quality Attribute for Biopharmaceuticals

Hendel JL, Kozak RP, Morgan CL, Royle L
Presented at: WCBP 2018: 22nd Symposium on the Interface of Regulatory and Analytical Sciences for Biotechnology Health Products
Washington D.C., United States. January 30-February 1st 2018





WCBP Sialic Acid Poster

Analysis of Sialic Acids in Biopharmaceuticals

Hendel J, Royle L, Kozak RP, Fernandes DL
Presented at: WCBP 2017: 21st Symposium on the Interface of Regulatory and Analytical Sciences for Biotechnology Health Products
Washington D.C., United States. January 24-26th 2017

Keywords: RP-HPLC, EPO, Mabs, fetuin, sialic acid, critical quality attribute (CQA), quantitation, Neu5Ac, Neu5Gc, DMB






Ludger Sofarchi 2016 poster

Automated High-Throughput Permethylation for Glycosylation Analysis Using MALDI-TOF-MS

Shubhakar A, Spencer D, Wuhrer M, Fernandes DL
Presented at the 38th Annual Congress of Pharmacology Society of Chile
Castro, Chile. Nov 26-29th 2016

Keywords: Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS), permethylation, high throughput (HTP), N-glycans, O-glycans





Ludger poster

Biopharmaceutical Erythropoietin Characterisation: critical quality attribute (CQA) mapping using LC-ESI-Qtof and automated database searching of glycopeptide analytes

Spencer D, Smith J, Gardner R, Galvin B, Resemann A, Hufnagel P, Fernandes D
Presented at the 64th ASMS Conference: American Society for Mass Spectrometry
San Antonio TX, United States. June 5-9th 2016

Keywords: erythropoietin (EPO), glycan, glycopeptide, LC-ESI-QTof-MS, ProteinScape, critical quality attribute (CQA)




Ludger poster

Procainamide labelling as part of a flexible glycoprofiling system for monitoring of Gal-a1-3Gal related Glycosylation Critical Quality Attributes (GCQAs) of monoclonal antibody (mAb) therapeutics throughout the product life cycle

Kozak RP, Royle L, Liew LP, Spencer DI, Fernandes DL
Presented at WCBP 2016: 20th Symposium on the Interface of Regulatory and Analytical Sciences for Biotechnology Health Products
Washington DC, United States. January 2016

Keywords: Gal alpha 1-3 gal, IgG, monoclonal antibody (mAb), glycosylation critical quality attribute (GCQA), QbD, procainamide, UHPLC, ESI-MS/MS




Ludger poster

A QbD-compatible approach for reliable measurement of sialic acid O-acetylation as a potential Glycosylation Critical Quality Attribute (GCQA) of erythropoietin (EPO) therapeutics

Fernandes DL, Thomson R, Gardner R, Liew LP, Kozak RP, Royle L, Strohfeldt K, Osborn H, Spencer DI
Presented at WCBP 2016: 20th Symposium on the Interface of Regulatory and Analytical Sciences for Biotechnology Health Products
Washington DC, United States. January 2016

Keywords: QbD, erythropoietin (EPO), sialate O-acetyl esterase (Nan S), glycosylation critical quality attribute (GCQA), sialic acids, DMB, procainamide, UHPLC, LC-MS




Ludger poster

A Fluorescent Labeling and Enrichment System for Glycopeptides Generated from Proteolytic Digestion of IgG mAbs;
A System That Can Be Used as Part of the Peptide Mapping Workflow

Hendel JL, Badia-Tortosa C, Spencer DI, Fernandes DL
Presented at USP: Glycosylation Analysis for Biopharmaceuticals Workshop
Washington DC, United States. August 2015

Keywords: glycopeptide, IgG, monoclonal antibody (mAb), peptide mapping, V-Tag, UHPLC, MALDI-MS




Ludger poster

Reliability of a semi-automated, high throughput (HT) MS based glycomics system for discovery of glycan biomarkers and Quality by Design (QbD) studies of glycoprotein therapeutics

Shubhakar A, Spencer DI, Wuhrer M, Fernandes DL
Presented at Gordon Research Conference
Lucca, Italy. March 2015




Ludger poster

Drug Glycosylation Analysis using Automated,
High Throughput, Orthogonal Methods

Shubhakar A, Kozak RP, Badia-Tortosa C, Gardner RA, Royle L, Wuhrer M, Spencer DI, Fernandes DL
Presented at PEGS 2014: Glycosylation Analysis for Biopharmaceuticals Workshop
Boston, United States. May 2014

Keywords: high throughput, permethylation, V-Tag, glycosylation critical quality attribute (GCQA), IgG, erythropeitin (EPO), UHPLC, MALDI-MS




Ludger poster

Optimisation of drug glycosylation supported by an automated high throughput permethylation MALDI-MS method

Shubhakar A, Spencer DI, Kozak RP, Velez-Suberbie L, Bracewell DG, Fernandes DL, Wuhrer M
Presented at ICS: 27th International Carbohydrate Symposium
Bangalore, India. January 2014

Keywords: high throughput, glycosylation critical quality attribute (GCQA), IgG4, bioreactor, permethylation, UHPLC, MALDI-MS




Ludger poster

Systematic Analysis of Drug Glycosylation Critical Quality Attributes (GCQAs)

Royle L, Kozak RP, Liew LP, Emery RJ, Fernandes DL
Presented at Glyco22: 22nd International Symposium on Glycoconjugates
Dalian, China. June 2013










Ludger poster

Suppression of Peeling during the Release of O-glycans by Hydrazinolysis

Kozak RP, Royle L, Gardner RA, Fernandes DL, Wuhrer M
Presented at the 4th Warren Workshop
Athens, Georgia, United States. August 2012










Contacts

R&D Programmes

Dr. Daniel Spencer

Dr. Daniel Spencer
Head of Development
daniel.spencer@ludger.com