demonstrating comparability and consistency of biologics ......demonstrating comparability and...

Demonstrating Comparability and Consistency Demonstrating Comparability and Consistency of Biologics Using LC/MS in Compliant

Laboratories

Patrick BoyceBiopharmaceutical Marketing Manager

Europe & IndiaWatersWaters

©2011 Waters Corporation

Overview

Challenges for LC/MS in compliant environments

Biopharmaceutical System Solution with UNIFIBiopharmaceutical System Solution with UNIFI

— Software architecture

— Compliance tools

— Methods for comparability & consistency


Characterization Characterization –– ICH Q6B ICH Q6B GuidelinesGuidelines

Extensive characterization of a biotechnological or Extensive characterization of a biotechnological or biological product is necessary to allow relevant specifications to be established

New analytical technology should be utilized when New analytical technology should be utilized when appropriate

Characterise primary structure, and higher order structure as possible

Define pattern of heterogeneity & demonstrate consistency

Product related substances should be characterized

Product & process related impurities should be characterised

©2011 Waters Corporation Company Confidential 3

BiosimilarsBiosimilars ––characterization & comparabilitycharacterization & comparabilityEMEA/CHMP/BWP/49348/2005EMEA/CHMP/BWP/49348/2005/ / / // / / /

“an extensive comparability exercise will be required to demonstrate that the similar biological medicinal to demonstrate that the similar biological medicinal product has a similar profile in terms of quality, safety and efficacy to the reference medicinal product”

“Extensive state of the art characterisation studies Extensive state-of-the-art characterisation studies should be applied to the similar biological and reference medicinal products in parallel at both the active substance and the medicinal product levels to demonstrate substance and the medicinal product levels to demonstrate with a high level of assurance that the quality of the similar biological medicinal product is

bl t th f di i l d t “comparable to the reference medicinal product. “


Biotherapeutic Protein LC/MS Applications

LCMS intact protein analysesMS t d t— MS exact mass data

LCMS peptide mapping analysesp p pp g y— MS and MSE exact mass data

Other Common Optical and MS Data workflowsOther Common Optical and MS Data workflows— SEC UV and SEC-MS analysis

— IEX UV analysis

— RP analysis


Some challenges for analytics in Some challenges for analytics in compliant organizationscompliant organizations

Electronic records

p gp g

— EU GMP Annex 11 and FDA CFR 21 Part 11

o Secure data with controls to restrict access

o Audit trailso ud t t a s

o Computers systems validation

Validation of analytical proceduresValidation of analytical procedures— ICH Q2

o Accuracy, precision, selectivity, detection limit, ifi i li i li i b quantification limit, linearity, range, robustness as

appropriate


Analysis times &, especially for biotherapeutics, resolution

Traceability -Linking Information to Recordsg

Standards usedfor Calibration

Samplefor CalibrationSets

Unchanged

Original Processing Method

CalibrationCurves

UniqueUniqueAnalysisAnalysis

Unchanged Raw Data File

Who Collected Wh P d

AnalysisAnalysisOriginal Instrument Method


Who ProcessedWho ReviewedWho Approved

LC/GC System Used

Product Code/Stage ReagentLIMS ID

E-cord information

Some challenges for high resolution LC/MS Some challenges for high resolution LC/MS analytics in compliant organizationsanalytics in compliant organizations

Secure data storage

y p gy p g

Secure data storage— Most high resolution MS acquisition software runs on

workstations only

Maintaining traceability— Most existing intact protein & peptide map LC/MS workflows

involve acquisition, followed by export of data to other software packages to perform specialized data processing and to create reports

S hi h l ti MS i iti ft d d t — Some high resolution MS acquisition software and data processing software packages have limited compliance tools for traceability and change control


Workflow for Routine Intact Protein and Workflow for Routine Intact Protein and Peptide Map Analysis by LC/MSPeptide Map Analysis by LC/MS

Import data into bioinformatics

Create Acquisition

Data Acquisition

bioinformatics software

Acquisition Method

and Sample Sequence

Data Acquisition

Who did what, when

Reports Compiled

and why?

Are my system

PrepareStandards & S l


and data

fit for purpose?

& Samples

Data Management

Overview

Challenges for LC/MS in compliant environments

Biopharmaceutical System Solution with UNIFIBiopharmaceutical System Solution with UNIFI

— Software architecture

— Compliance tools

— Methods for comparability & consistency


The Waters Biopharmaceutical The Waters Biopharmaceutical System SolutionSystem SolutionSystem SolutionSystem Solution

An analytical platform for UPLC/UV and UPLC/MS biotherapeutic analysis deployable across unregulated and GxP laboratories

XEVO G2 Tof(MS and MSE)

TUV Optical Detector

ACQUITY UPLC ACQUITY UPLC H-Class BIO


UNIFI Scientific Information SystemBioSeparations Chemistries

Performance and Usability through Engineered Simplicityg g p y

Automatically ensuring the system is ready to run Automatically ensuring the system is ready to run


The Most Extensive Range The Most Extensive Range of Interface Capabilitiesof Interface Capabilitiespp

Universal source architecture Enables the widest range of ionization

techniques to be utilized

ESI APCI ESCIESI, APCI, ESCI

nanoFlow ESI, TRIZAIC

Simple and tool-free All i t h b t

APPI, ASAP, APGC

Compatible with third party sources


Allowing you to change between ion source options in minutes.

p p y(DESI, DART, etc)

QuanTof Technology for Stability of QuanTof Technology for Stability of Mass Accuracy over TimeMass Accuracy over Timeyy

M d M A f P tid

10

Measured Mass Accuracy for Peptide THTCPPCPAPELLGGPSVFLFPPKPK in 96 hrs

2

6

(ppm

)

-2

2

0 5 10 15 20 25 30

Mas

s Er

ror (

-10

-6

M


10# of Injections

ACQUITY UPLC HACQUITY UPLC H--Class BioClass BioQQ

Ultimate chromatographic performanceL di i t b 2 l—Low dispersion system sub-2 µm columns

Biocompatible system—Eliminate system corrosionEliminate system corrosion—Best sample recovery

Flexibility, robustnessl bl d—Quarternary solvent blending

—Suitable for RP, IEX, SEC, HILIC—Robustness tested for high salt—AutoBlend+ for dial-up buffers

Needle in flow path designLow carryover performance


—Low carryover performance

Peptide MappingPeptide MappingReversedReversed--phase chromatographyphase chromatographyp g p yp g p y

The peptide mapping sample list started on Friday and ran automatically th h th k d Th d t d t i t t k


through the weekend. The data was ready to review upon return to work on Monday. Every third sample is shown, demonstrating excellent reproducibility in both resolution and retention

Transfer Methods With EaseTransfer Methods With Ease

Utilize existing Assets

Transfer from UPLC-to-HPLC

Future-proof your lab

Run HPLC methods on ACQUITY UPLC H-Class


BioseparationBioseparation ChemistriesChemistriesBEH™ Technology ParticlesBEH™ Technology ParticlesBridged Bridged EthylSiloxaneEthylSiloxane/Silica Hybrid/Silica HybridBridged Bridged EthylSiloxaneEthylSiloxane/Silica Hybrid/Silica Hybrid

Si

EtO CH2 CH2

Si OOEt

SiOEt

O EtEtO OEtEtO

O

Si

EtO

Si O

O

OEtO

O

Si OOEt

Et

+ SiEtO

EtOCH2

EtO

CH2Si

OEt

OEtOEt

4 SiEtO

EtO OEtEtO

1

Polyethoxysilane(BPEOS)

OEt nTetraethoxysilane(TEOS)

Bis(triethoxysilyl)ethane(BTEE)

Bridged EthanesIn Silica MatrixIn Silica Matrix


Anal. Chem. 2003, 75, 6781-6788

U.S. Patent No. 6,686,035 B2

BioseparationBioseparation Application Tested Application Tested ChemistriesChemistries

Protein Separation Technology— RP, SEC, IEX

Peptide Separation Technology— RPRP, SEC, IEX RP

Oligonucleotide Separation Technology— RP

Glycan Separation Technology— RP, SEC, IEX


Built on decades of analytical data and informatics experience.p


KEY FEATURES: Solution Architecture

One software for LC with UV and MS detection

Robust Set of Data Analysis and Reporting Tools

Fully Configurable GxP Compliance Tools— Named user login and authentication— Electronic signatures— Audit trails and Action-Explanation Policies— Electronic System Qualification and Maintainance Tools

Future-proof: Workstation to Workgroup to Enterprise scalability


UNIFI Workstation Architecture

One server handles both functions:

Arch

itect

ure

• Low-level (Database management and Instrument Control)

• High-level (Data processing and

Data Processing

Data/Results Browser

Database (Data/Results)

Wor

ksta

tion

A

g ( p gResults browser)

Typically a workstation is physically Instrument Control (ISS)

Typically a workstation is physically located next to the LC/MS system.

Lenovo D20 WorkstationXeon E5504 2.0 GHz processor 12 GB GDDR3 RAM Nvidia Quadro FX 1800 graphics card


Nvidia Quadro FX 1800 graphics card Windows 7 64-bit operating system

Achieving a Regulatory Compliant-ready deployment of UNIFIy p y

UNIFI capabilities will be extended from the workstationimplementation to the compliant-ready architecture now used for

t k d Ch D t S t ( W t E CDS)

Data/Results Browser Data/Results BrowserData/Results BrowserData/Results Browser Data/Results BrowserData/Results Browser

networked Chrom Data Systems (e.g. Waters Empower CDS).

Client PC Client PCClient PCClient PCClient PC Client PCClient PCClient PC

Data Server

LaboratoryNetwork

CompanyIntranet

Instrument Data Server

LaboratoryNetwork

CompanyIntranet

Instrument

Data Processing Apps


I t t C t l

Systems ServerData Processing Apps


I t t C t l

Systems Server


Instrument ControlInstrument Control

UNIFI Scientific Information Systemy

Automates routine data processingAutomates routine data processing

Task-based, workflow oriented design, g

Intuitive interface tailored to users’ roles and permissions

A method covers acquisition, processing & bioinformatics, reporting, and electronic signatures

f l d d d b l


Powerful SDMS-derived data mining capabilities

UNIFI Tools for Compliance p

Named User Authentication User role(s) User role(s) — Limit capabilities within system/workflows— Limit access to data/information

A dit t ail of se acti ities

Who did what, h d Audit trail of user activities

User action explanation policies (configurable)Electronic Signatures (21CFR11)

when and why?

Software IQ/OQAre my system So t a e Q/OQ

System IQ/OQ/PQ (Electronic Documentation)

System Maintenance (Electronic Documentation)

system

and data

fit for


Data Validation via SHA1 Checksums fit for purpose?

Basic OQ and Extended OQ Basic OQ and Extended OQ vsvs PQPQQ QQ Q QQ

IQ Simple PQ or UAT’sIQ OQ PQ or UAT s

Simple IQ Simple OQPQ or UAT’s PQ or UAT’sVendor Factory Testing

For many years this was sufficient

Larger companies through CSV validation groups, C lt t d GAMP d ti i

IQ Extended OQ PQ or UAT’s

Consultants and GAMP recommendations now increase the PQ to a much higher level and full lifecycle validation needed.IQ

Comprehensive Validation Service PQ and full Docs ( VP and VR)


Significantly increases the Validation Effort

Instrument QualificationInstrument Qualification

Built-in Qualification and Maintenance Centre stores:

QQ

Built in Qualification and Maintenance Centre stores:— Qualification protocols

— Engineers’ certification

Qualification reports— Qualification reports

— Qualification results

— Maintenance log


Methods cover data acquisition to electronic sign-off of a report

Systems andData

Acquisition

Data P iProcessing

andBioinformatics


Automated Routine Data Processingg


Automated comparative analysis Automated comparative analysis ––peptide map analysispeptide map analysisp p p yp p p y

Common and unique peptides identified

Coverage maps for reference and reference and

analyte

h f


Chromatograms for reference (top) & analyte (bottom)

Automated comparative analysis Automated comparative analysis ––intact protein analysisintact protein analysisp yp y

Protein modifications identified

h f


Deconvoluted annotated mass

spectra

Chromatograms for reference and

analyte

Applying a Reporting TemplateApplying a Reporting Template

Report Output


Report Output

Trending of results for larger data sets


Trending of results for larger data setsdata sets

Component B – Modified Form

Trend of intensity: Specific components tracked –e.g. a check for modification out of spec, traceable to the batch; or studies of variation in an attribute

Component A – Unmodified Form

to the batch; or studies of variation in an attribute (glycoforms)


Blanks

Simple Intact Antibody LCMS Analysis Reporty p



Import data into bioinformatics

Create Acquisition

Data Acquisition

bioinformatics software

Acquisition Method

and Sample Sequence

Data Acquisition

Who did what, when

Reports Compiled

and why?

Are my system



and data

fit for purpose?

& Samples

Data Management


Data AcquisitionData Acquisition& ProcessingCreate Analysis Method

and Sample Sequence

Reports Compiled, Reviewed & Signed



& Samples

Data Management

Relevant literature downloadsRelevant literature downloads

Search www.waters.com for:

— Biopharmaceutical System Solution with UNIFI

— Biopharmaceutical System Solution with UNIFI White Paper

UNIFI S i tifi I f ti S t— UNIFI Scientific Information System

— Biopharmaceutical System Solution with UNIFI: Intact Protein Characterization

— Biopharmaceutical System Solution with UNIFI: Simplifying Verification of Peptide Mapping Results

— Xevo G2 Tof Brochure

— BioSeparation Products and Columns


Summary

Deploying LC with high resolution MS in compliant organizations presents challengesorganizations presents challenges

Purpose-built technologies for Biotherapeutic analysis

— Secure data

— Traceable results

— Acquisition, automated data processing, comparative analysis, trending and report generation all within one softwaretrending and report generation all within one software


demonstrating comparability and consistency of biologics ......demonstrating comparability and...

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