Analysis of Protein Biopharmaceuticals

Solviasprovidescomprehensivephysicalchemistryservices for the identificationandselectionof

polymorphs,salts,andco-crystals,includingmethoddevelopmentforpolymorphicpurityandthe

optimization of crystallization processes. With years of drug development experience as a former

researchgroupofamajorpharmaceuticalcompany,wecanhelpyoucrystallizeyourideas.

Solvias AGP.O.Box

4002BaselSwitzerland

Tel. +41616866161 Fax+41616866565

[email protected]

Analysis of Protein Biopharmaceuticals

Comprehensive cGMP Services at Every Stage of Drug Development

At Solvias, we work closely with you …

Amazing where you can go

Primary activities

• Characterization(forregulatorysubmission)

• Methoddevelopmentandvalidation(ICH)

• QCReleasetesting(cGMP)

• Stabilitystudies(cGMP)• Comparability

Additional services include

• Certification,storageandsupplyof

customer-specificreferencesubstances

• Analyticalsupportinprocessvalidation

• Analyticalsupportinformulation

development

• Extractablesandleachables

• Analyticaltroubleshooting

Quality

• FDA-inspected

• cGMPcontractlaboratoryapprovedby

Swissmedic

• ISO9001-certifiedQMsystem

ServicesBioanalytical programs individually tailored to meet your needs

Solviasprovidescomprehensiveservicesforpro-

tein-baseddrugs,tobiotechnologyandpharma-

ceuticalcompanies,ateverystageofdrugdeve-

lopment.

… to design customized programs …

Solvias can help you to solve your most com-

plex analytical challenges by providing expert

guidance and by designing flexible and custo-

mizedprograms.Complexprojectsarecoordina-

ted by dedicated professional customer project

managers.

… that meet your needs …

Methods destined for use in pharmaceutical

release testing are developed with a focus on

robustness and reproducibility. The goal of the

methoddevelopmentprogramistoestablish

a reliable method and carefully describe it in a

standard operating procedure (SOP) that can

readily be executed at Solvias or transferred to

yourlaboratoryofchoice.Withintheframework

of the method development program, critical

parameters such as the linearity, reproducibi-

lity, and LOQ will be checked to ensure reliable

analytical results. A written SOP that includesproductspecificationsisaprerequisiteforbegin-

ningamethodvalidationprogram.Themethod

validation program is performed under cGMP

according to ICH guidelines and typically in-

cludes the following parameters: specificity and

selectivity,accuracy andprecision,limitofdetec-

tion(LOD)/limitofquantification(LOQ),linearity

andmeasurementrange,robustness,andsoluti-

onstability.

A milestone-based program can easily be de-

signedandadaptedtobestfittheneedsofyour

drugdevelopmentprogram.Allproduct-specific

intellectualproperty(IP)formethodsdeveloped

bySolviasisassignedtothecustomer.

… and are tailored to your drug.

The selection of the techniques to be used is

baseduponthepropertiesofthedrug.Forexa-

mple, toestablishastability-indicatingmethod,

techniques such as CE and HPLC are applied to

samples that have been stressed (e.g. tempe-

rature). As both are orthogonal methods with

different separation processes, they reveal a

complementarypictureofproduct-relateddegra-

dation forms. The optimal method can then be

furtherdeveloped.Wepreferentiallyapplyquan-

titative methods, if quantification is desired

(e.g.capillaryelectrophoresisinsteadofflatbed

electrophoresis).

Solvias AGP.O.Box

Tel. +41616866161 Fax+41616866565

[email protected]

Primary activities

development

Quality

• FDA-inspected

Swissmedic

lopment.

managers.

onstability.

electrophoresis).

Disulfide bridging

• Enzymaticcleavageofthenativeprotein

LC-MSseparationandidentificationofpeptide

fragments

• Proofofproperfoldingbypresenceofspecies

withproperdisulfidebridgingandabsence

ofimproperlybridgedfragments

• Reductivealkylationtoinduceshiftofbridged

peptidesinthechromatogramasacontrolstep

Carbohydrate analysis in release testing

• SialicacidbyHPLC

• NeutralsugarsbyHPLC

• Glycosylationwithandwithoutsialicacid

bycapillaryelectrophoresis

• HPAEC-PAD

Biosimilar/Follow-on Biologics

• Comprehensivechoiceofstate-of-the-art

methodology

• Experiencetoensuresuccessfulregistration

• Dedicatedprojectmanagement

Technology baseA broad range of capabilities allows Solvias to apply the best solution to your problem

We focus on reliable and efficient methods for pharmaceutical release testing

• Capillaryelectrophoresis(CZE,CE-IEF,CE-SDS;

UVandLIFdetection)

• Chromatography(HPLC,GC,DC,SEC,IEC;

manyspecialdetectors)

• Electrophoresis(IEF,SDS-PAGE,nativegel;

standardizedandreadygels)

• Aminoacidanalysis

• Massspectrometry(ESI,MALDI-TOF/TOF-MS)

• Westernblotting

• ELISA,ILA

• Hyphenatedtechniques(LC-MS)

• QuantitativePCR,thresholdsystem

• Spectroscopy(UV/VIS,CD,fluorescence)

• Analyticalultracentrifugation

• Lightscattering(MALS,DLS)

• DNAsequencing(cGMP)

ApplicationsComplete characterization programs according to ICH Guideline Q6B: comprehensiveness is our strength

• PeptidemappingbyLC-MS

• DisulfidebridgingbyLC-MS

• Determinationofoxidativeforms

bye.g.LC-MSandLC-UV

• Isoelectricfocusingbycapillary

electrophoresis

• PEGsubstitutionbycapillaryelectrophoresis

andmassspectrometry

• Carbohydrateanalysisbye.g.HPLC

andcapillaryelectrophoresis

HPAEC-Dionex,MALDI-TOF/TOF

massspectrometry

• Contentdeterminationbyaminoacidanalysis

• Determinationofextinctioncoefficientby

aminoacidanalysisandUV/VISabsorbance

• QuantificationofTween®

Practical solutions to complex problemsSolvias brings years of pharmaceutical experience to solve the most complex analytical and regulatory challenges

Cf.ourpublication:Applicationofcapillaryzoneelectrophoresisandreversed-phasehigh-performanceliquidchromatographyinthe

biopharmaceuticalindustryforthequantitativeanalysisofthemonosaccharidesreleasedfromahighlyglycosylatedtherapeuticprotein,

K.Racaityté,S.KiessigandF.Kálmán(SolviasAG):

Journal of Chromatography A,Volume1079,Issues1–2,24June2005,pages354–365.

Separation of protein pl markers by CE-IEF

Plot of the isoelectric point versus migration time obtained by CE-IEF

Structuralcharacterization• Aminoacidsequence• Aminoacidcomposition• Terminalaminoacidsequence• Peptidemap• Sulfhydrylgroupsanddisulfidebridges• Monosacchorideanalysis• Carbohydratestructure• Spectroscopy

Physicochemicalproperties• Molecularweight• Molecularsize• Isoformpattern• Isoelectricpoint• Extinctioncoefficient• Electrophoreticpatterns• Liquidchromatographicpatterns• Spectralproperties• Glycananalysis• PEGanalysis

Quantity• A280

• Quantitativeaminoacidanalysis• Nitrogendetermination• Proteinassay(Lowry)• Immunoassay(ELISA)

• Buffer/matrixcomposition(anion,cation,Tween®)

• Residualwater(KarlFischer)

Product-relatedimpurities• Degradation• Aggregation(dimers,higher oligomersandaggregates)• Proteinoxidation• Proteindeamidation• Disulfidescrambling

Contaminants• Microbialcontamination• Endotoxin• Viruscontamination• Heavymetals

Process-relatedimpurities• Residualsolvents• Leachables

(e.g.ligandsforaffinitychromatography)• Cellmediacomponents

(e.g.growthhormones)• ResidualDNA• Residualhostcellprotein• Detergents

Purity• Determinationofmolecularentities

composingthedrugsubstance

Expipiants• Allclassesofmolecule

Disulfide bridging

fragments

• HPAEC-PAD

methodology

UVandLIFdetection)

• Westernblotting

• ELISA,ILA

bye.g.LC-MSandLC-UV

electrophoresis

andmassspectrometry

massspectrometry

Quantity• A280

Disulfide bridging

fragments

• HPAEC-PAD

methodology

UVandLIFdetection)

• Westernblotting

• ELISA,ILA

bye.g.LC-MSandLC-UV

electrophoresis

andmassspectrometry

massspectrometry

Quantity• A280

Disulfide bridging

fragments

• HPAEC-PAD

methodology

UVandLIFdetection)

• Westernblotting

• ELISA,ILA

bye.g.LC-MSandLC-UV

electrophoresis

andmassspectrometry

massspectrometry

Quantity• A280

Solvias AGP.O.Box

Tel. +41616866161 Fax+41616866565

[email protected]

Primary activities

development

Quality

• FDA-inspected

Swissmedic

lopment.

managers.

onstability.

electrophoresis).

Solvias provides comprehensive physical chemistry services for the identification and selection of

polymorphs, salts, and co-crystals, including method development for polymorphic purity and the

research group of a major pharmaceutical company, we can help you crystallize your ideas.

Solvias AGRömerpark 2

4303 KaiseraugstSwitzerland

Tel. +41 61 845 60 00 Fax +41 61 845 69 00

[email protected]

Primary activities

• Characterization (for regulatory submission)

• Method development and validation (ICH)

• QC Release testing (cGMP)

• Stability studies (cGMP)

• Comparability

• Certification, storage and supply of

customer-specific reference substances

• Analytical support in process validation

• Analytical support in formulation

development

• Extractables and leachables

• Analytical troubleshooting

Quality

• FDA-inspected

• cGMP contract laboratory approved by

Swissmedic

• ISO 9001-certified QM system

Solvias provides comprehensive serv ices for pro-

tein-based drugs, to bio techno l ogy and pharma-

ceutical companies, at every stage of drug deve-

lopment.

mized programs. Complex projects are coordina-

managers.

method development program is to establish

your laboratory of choice. Within the framework

an alytical results. A written SOP that includes

product specifications is a prerequisite for begin-

ning a method validation program. The method

selectivity, accuracy and precision, limit of detec-

tion (LOD) / limit of quantification (LOQ), linearity

and measurement range, robustness, and soluti-

on stability.

signed and adapted to best fit the needs of your

drug development program. All product-specific

intellectual property (IP) for methods developed

by Solvias is assigned to the customer.

based upon the properties of the drug. For exa-

mple, to establish a stability-indicating method,

complementary picture of product-related de gra-

further developed. We preferentially apply quan-

(e.g. capillary electrophoresis instead of flat bed

electrophoresis).

Analysis of Protein Biopharmaceuticals

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