GenScript Your Biology Research Partner · 2019. 6. 11. · GenScript, A Global Biology CRO. Global headquarters Piscataway, New Jersey, USA. Main production site Local technical

GenScriptYour Biology Research Partner

September 28, 2017

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Company IntroductionMolecular Biology ServicesProtein ServicesAntibody ServicesPeptide ServicesADD Services


Intellectual Property Protection


GenScript, A Global Biology CRO

Global headquartersPiscataway, New Jersey, USA

Main production siteNanjing, China Local technical support

24-hour customer service Fast global logistics Competitive pricing Stringent IP protection

Founded in 2002 Publicly traded at Hongkong Stock Exchange

(HKG:01548) 1300+ employees No. 1 in gene synthesis No. 1 in Bio-CRO in China Offering one-stop reagent production and

discovery services


CRO Leadership

Quality Awards Innovation Awards Productivity Awards Reliability Awards


Services Overview

GenScriptProducts & Services

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Gene

Peptide

Protein

Antibody

Antibody Drug

Discovery

Cell Line

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Molecular Biology Services

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Molecular Biology Capacity Overview

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Successful delivery of more than 600,000 genes, success rate 99.95% Largest capacity at 100 million bp/month

First commercial partner invited to join Johns Hopkins University Yeast Genome Project (Sc2.0); Collaborate with Macquarie University (Chr. 14 & 16)

Partnered with Novovax to synthesize H7N9 genes within 6 days to enable development of a vaccine within 6 weeks of the first reports of this virus in humans.


MB Related Services

De novo Gene Synthesis

Cloning

Mutagenesis

Plasmid Preparation

GenEZ ORF Clone

CloneArk Clones

GenScript GenEZ ORF database:• 40,000 in-stock Human/Mouse/Rat

ORFs• Over 10,000 clone in default vector

for next-day shipping• Over 2 million from 186 different

species could be synthesized

GenScript Gene Synthesis:• Any DNA sequence, even complex,

GC-rich, repeated, or long genes, with 100% sequence accuracy guaranteed

• Free codon optimization • Guaranteed sequence

GenScript CloneArk database:• Details of all gene constructs you’ve

ever ordered from GenScript• Easy record-keeping• Convenience any time to order new

plasmid preps, mutant or other service

GenScript Cloning Service:• Ready-to-use construct – not a

semi-finished part• Unbeatable price of only $49• 2-day turnaround added to your

gene synthesis order

GenScript Mutant Service:• Introduce mutations at any site• Mutations on Large DNA Constructs• Most Competitive Prices starting at

$99/mutant

GenScript Plasmid Service:• Microgram to gram production• Extremely low levels of endotoxin:

≤0.01 EU/µg• Minimum of 7 stringent QC

parameters analyzed for each step• Only an additional 3 days with any

gene synthesis order

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CRISPR Service Portfolio and Resources

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Designed by Feng Zhang’s laboratory at Broad Institute

Over 2,000 gRNA constructs ready to delivery

Pre-designed crRNA and tracrRNA bundled with cas9 protein to improve specificity

GeCKO, SAM library and Pathway-focused library are provided

GenCRISPR™produces a genetically modified cell using any mammalian cell line and targeting any gene

GenScript offers validated CRISPR products, services and resources to help you harness the power of CRISPR genome editing for your research.

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Protein Services

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Protein Capacity Overview

Strong Track Record and Global Presence >6,000 batches of high quality proteins delivered >1,500 batches of high quality rAbs delivered >92% success rate for all projects Successfully delivered a variety of protein targets


Expression Systems

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Transient Expression of Recombinant Protein & Antibody• Fast production and delivery of high quality• Gene to rAb as soon as 4 weeks，gene to protein as soon as 6 weeks• From pilot scale to guaranteed production, up to gram quantities.

Stable Cell Line Development

Process Development GLP-Compliant Bioprocess

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Mammalian Expression

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BacPower™ Guaranteed Bacterial Protein Expression Services

BacPower™ Customized Services

Fermentation services(5L-500L, Batch/Fed-batch)

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Bacterial Expression

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Antibody Services

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Antibody Capacity Overview

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Global headquartersPiscataway, New Jersey, USA

AAALAC International and OLAW accredited animal facility• 6,000 sq.m, hosting more than 8,000 rabbits and 12,000 rodents• Providing a broad range of antibody services from individual cases to

bulk orders

High volume output

Monoclonal Antibody Project 50-80/Month

Polyclonal Antibody Project 500-600/Month

Antibody Related Project 50/Month


Antibody Services Introduction

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o 12+ years experience, delivered 200+ anti-ID orders with 100% success rateo High Specificity

Use isotype (usually Human IgG1 and IgG4) counter screen to remove unspecific cloneso High affinityMouse anti-ID mAb (in nM range), rabbit mAb service for higher affinity o Two different anti-ID to detect both free drug and total drug

Antigen-blocking (neutralizing) anti-IDs detect free drug Non-blocking (non-inhibitory) anti-IDs detect free and bound drug

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Why GenScript anti-ID?

Rabbit polyclonal Anti-ID

Rabbit monoclonal Anti-ID

Mice monoclonal Anti-ID


Anti-ID services Details

Services Anti-ID monoclonal antibody Anti-ID polyclonal antibody

Starting material Target antibody drug 2-3mg Target antibody drug 20 mg or more

Work flow

QC Cross-reactivity with control IgG <10% Cross-reactivity with control IgG <10%

Deliverables Hybridoma cell lines, supernatants and purified anti-ID antibody (optional) 0.5-3mg purified anti-ID antibody/rabbit


GenScript rabbit mAb technology

o Hybridoma technology for rabbit mAb generation• Experienced in hybridoma generation• A well-developed hybridoma platform (more cost-efficient compared with others)

o GenScript’s hybridoma technology• Rabbit-Mouse (Rb-Ms) chimeric hybridoma technology, without IP issues• Overcome the technical difficulties of Rb-Ms hybridoma through two strategies:

1) Improving the fusion efficiency and hybridoma stability2) Combining antibody sequencing technology at an earlier stage

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Peptide Services

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Microwave technology: Efficient, high-yield peptide synthesis AccuPep+ QC guarantee high quality peptides Ligation Technology: Long peptide synthesis (up to 200 AA) Over 10,000 peptides delivered per month Over 300 modifications Advantages:

• Guaranteed delivery• High-efficiency• Improved peptide yields• Decreased cost• Peptides of any length• Easier modifications

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Peptide Capacity Overview


Peptide Services Introduction

Custom Peptide Synthesis

Rush Peptide Synthesis

Fast Peptide Synthesis

Sequence Length Up to 200 AA 5-10 AA 11-20 AA 5-20 AA

Purity All purities Crude Crude >70% to >95%

Quantity Mg to kg < 20 mg < 20 mg

Modifications Comprehensive(over 300

modifications)

Amidation (C-terminal)Acetylation (N-

terminal)

Amidation (C-terminal)Acetylation (N-terminal)

Starting Turnaround Time

2-3 weeks 5 days 7 days 9 days

Custom peptides service Flexible quantity, purity peptides with economical prices

Express peptide service Fast peptides & Rush peptides, get your peptides from just 5 days!

Peptide library & Peptide array service Standard library & micro-scale library with different quantities for your drug and

vaccine development. Quick and cost-efficient custom peptide array service

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Antibody Drug Discovery

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Your Partner from Research to IND

Discovery

Development

Testing

Lead Optimization Humanization Affinity maturation Developability assessment

Lead GenerationAntigen preparation ImmunizationCell fusion and HTP screeningSubcloning and stabilizationAntibody production & characterization

Biochemical & biophysical assayCell-based assayProtein analytics In vivo efficacy evaluationStability Studies

Transient ExpressionCell line developmentCell Culture developmentPurification developmentFormulation development

GMP scale-upTechnical transfer

IND


Discovery Roadmap

Discovery

Development

Testing

Milestone Timeline (months)1 2 3 4 5 6 7 8 9 10 11 12 13

1. Material PreparationProtein antigen, Overexpressing cell line for membrane target, Reference Ab,DNA plasmids for DNA immunization

2. ImmunizationProtein immunization for soluble targets and ECD of TM targets, Cell and DNA immunization for TM targets, ImmunoPlus modification for high homology targets, Carrier protein conjugation to enhance immune responses

3. Fusion & ScreeningCell fusions with ELISA and/or FACS screening, Other screening(such as ligand blocking screening, cross-species reactivity test)

4. SubcloningOne-round subcloning with ELISA and/or FACS screening, Final subcloning and cryopreservation of final subclones, Other screening (such as ligand blocking screening, cross-species reactivity test)

5. Antibody ProductionAntibody mini-production (0.1mg), Antibody midi-production (0.5-1mg),Antibody maxi-production (5-15mg)

6. Antibody CharacterizationELISA EC50, Binding by FACS for membrane targets, Species cross-reactivity test,ELISA IC50, Ligand blocking by FACS for membrane targets, Epitope binning,Affinity measurement, Antibody sequencing, In vitro functional assay

7. Antibody HumanizationChimeric antibody production and affinity confirmation, Humanization by rationale design, Thermal stability and expression level evaluation, Humanized antibody production and affinity determination


One-Stop Solution from Research to IND

Target Assessment & Validation

Ab Lead Generation

Ab Lead Characterization & Optimization

Candidate Production

& Characteri-

zation

GMP Manufacturing(with partner)

• Gene & Peptide synthesis• Antibody production• Protein production in

mammalian, E. coli, insect or yeast expression

• Cell line engineering

• Hybridoma development• Ab screening &

characterization200+ therapeutic antibody leads generation

• Ab sequencing• High Throughput Ab

production• Transient expression in

shaking flask/wave bag• Humanization• Affinity maturation50+ humanization projects delivered

• Stable cell line generation

• Process development• Small- to mid-scale

Production• Research Cell Banking

(RCB)• Bioanalytics & Analytics50+ cell line & process development projects delivered

• 20-1000L cGMP fermentation

12 mAb, 2 Fc-fusion protein, 1 r-protein, 4 protein vaccine, 2 plasmid DNA

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Accurate to the last base!

o Strict quality control ensures accurate sequencing resultso Track record of success: 3,000+ antibodies sequenced with 100% accuracy o Competitive pricing and turn-around time

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Antibody Sequencing


Timeline and Price:

Sample Requirements:

GenScript Sequencing Services Delivery Time Price Deliverable

Variable domain & leader sequence sequencing 1-2 weeks $1,200 Detailed work performance report; Sequence

data; Alignments of the heavy and light chains; Plasmid containing antibody sequence (optional)Full length antibody sequencing 2-3 weeks $2,000

Samples Quantity Shipping Requirements

Hybridoma cells (frozen, live, pellet) >1×10^6 cells Liquid nitrogen or dry ice

Antibody Sequencing Details

Candidate ProductionMammalian Transient & Stable Expression


Transient Expression

• HEK293 and CHO Cell Expression• Serum-free production• Rapid and high yield production• mg to gram-level Ab • Sequence to Ab in ≥5 weeks

Regular Transient Expression

• HD-HEK and HD-CHO expression• Serum-free production• Alternative approach for difficult proteins• Up to 3g/L Ab titer• 5-100 fold yield improvement• Sequence to Ab in ≥5 weeks

High Density (HD) Transient Expression

References:1. High-level and high-throughput recombinant protein production by transient transfection of suspension-growing human 293-EBNA1 cells. Durocher Y, Perret S, Kamen A.

Nucleic Acids Res. 2002 Jan 15;30(2):E9.2. Over-expression of secreted proteins from mammalian cell lines. Dalton AC, Barton WA. Protein Science : A Publication of the Protein Society. 2014;23(5):517-525.

doi:10.1002/pro.2439.


Stable Cell Pool

Use of GenScript Technology results in high-producing cell pools.

• Cell pools achieved within 8 weeks from point of transfection

• 400mg/L - 1400mg/L

Reference: Stable transfection pools for large quantity of protein production. Ye J. Methods Mol Biol. 2012;899:221-5. doi: 10.1007/978-1-61779-921-1_14


Stable Cell Line Development• Glutamine Synthetase based selection system• High productivities without amplification• Speed – no need for rounds of gene amplification• Scalable

GS

• CHO derived host cell line for expression• Adapted to suspension and AFCDM• Selected for superior growth and expression

CHO

• Fully optimized process• Product concentrations of >5g/L achieved• High probability of monoclonality

Cell Line Development

0

2

4

6

Pool Single cellclone

Batchculture

Fed-batchround 1

Fed-batchround 2

Tite

r (g/

L)

mAb Expression level

mAb Expression level

High volumetric productivity

Cost

GenScript has created cell lines producing >5g/L Ab titer with production rates in the range 50-100 pg/cell/day.

Reference: High-level expression of a recombinant antibody from myeloma cells using a glutamine synthetase gene as an amplifiable selectable marker. Bebbington CR, Renner G, Thomson S, King D, Abrams D, Yarranton GT (1992) Biotechnol. 10:169-175.


Protein Analytics

Parameter Analytical Test

Purity, Homogeneity SEC-HPLC,RP-HPLC, CE-SDS, SDS-PAGE

Primary Structure including deamidation, oxidation N-terminal sequencing, sequence coverage,

Higher Order Structure SEC-HPLC, CD, CE-SDS

Charge cIEF, IEC-HPLC

Size SDS-PAGE, SEC-HPLC

MW LC-MS

Hydrophobicity RP-HPLC, HIC-HPLC

Disulfide Bond Analysis LC-MS

Immunoreactivity and Function IP, Western Blot, Biochemical Assay

Glycosylation Profile HPLC,LC-MS

Safety Endotoxin Testing, HCP, HC DNA, Protein A residue

With LC-MS at its core, GenScript performs a variety of characterization testing to ensure integrity of the final product.


Services Overview

GenScriptProducts & Services

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Gene

Peptide

Protein

Antibody

Discovery Biology

Cell Line

Thank youExpertise, Flexibility, Solutions


BioProduction Cell Line Capability

GenScript USA, Inc and Horizon Discovery Group entered Bioproduction Cell Line Licensing agreement for the commercial use of CHO-Glutamine Synthetase cell line.

Widely accepted technology adopted by hundreds of companies and academic institutions and with several products in clinic (Zenapax®, Synagis®, Soliris®).

Flexible licensing with simplified terms.• For R&D 25,000 GBP/project ($33K)• For IND 150,000 GBP/ project ($200K)

No royalties. No IP burden.

GenScript has also partnered with global GMP-certified biologics developer and CMO to meet all your manufacturing needs.


Recombinant Ab Production Expertise

His

VL VH

VH

VL

ScFv1

ScFv2

BiScFv

Linker

C-Fc tag protein N-Fc tag protein

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Client • Start up Biotech Company.

Project

• Client has access to a proprietary technology for identifying and isolating antigen-specific B-cells.

• Engaged GenScript for production of a panel of anti-HIV Abs.

• Client focused on biology and developability of lead Abs to ensure efficacy while minimizing downstream risks.

Client Bottleneck

• Antibody Production of in-silico sequence optimized leads for HTP biophysical characterization studies.

Outcome

• Client submitted hundreds of sequences to GenScript.

• GenScript performed codon optimization, gene synthesis, cloning, plasmid prep, HTP transfection, expression and 1-step purification and delivered 1mg purified Abs as specified to client for their characterization studies.

• Further scale up for in vitro neutralization assays and in vivo protective activity assay in mouse and macaques.

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Case Study: HTP Antibody Production

Sequence to Antibody from $499 as fast as 6 weeks.

Ideal for Antibody Drug screening applications.

Ab Lead OptimizationHumanization & Affinity Maturation

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Antibody Humanization• Structure-based back mutation Library design and construction• FASEBA platform experimentally to boost efficiency• Over 50 projects delivered in past three years

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GenScript Humanization Services Delivery Time Price Deliverable

Express Humanization 2-3 months $41,000Detailed work performance report; Sequences of top 3

humanized antibodies

Detailed work performance report; Sequences of top 3 humanized antibodies; affinity guaranteedDeluxe Humanization 4-5 months

$80,000-100,000


Ligand Analyte ka (1/Ms) kd (1/s) KD (M) Rmax (RU) Chi² (RU²) U-value

WT-FabMembrane protein ECD

4.05E+05 7.14E-05 1.76E-10 394 13.2 7

Fab-BM1 4.50E+05 7.37E-05 1.64E-10 295.8 10.3 7

Fab-BM2 4.25E+05 6.82E-05 1.60E-10 455.7 18.4 7

Case study: Humanizing Antibody That Recognizes a membrane protein ECD

WT-Fab Fab-BM1 Fab-BM2

Fab-BM1: Humanized Fab containing back mutated residues IFab-BM2: Humanized Fab containing back mutated residues II

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Affinity Maturation

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Affinity maturation, to increase affinity by high-throughput screening approach

Identification of paratope, the essential step

Target, to reach at least low nM or improve 10-fold

5-6 months


Case Study: Affinity Maturation

Ligand ka (1/Ms) kd (1/s) KD (M) Rmax (RU) Chi² (RU²) Foldincrease

Wild-type IgG 6.35E+04 0.005986 9.43E-08 121.3 0.263 1AS04498 5.77E+04 1.44E-04 2.50E-09 129.4 0.336 38

Wild-type IgG

Ab Lead GenerationHybridoma Development


Murine Antibody Lead Generation

1o. In vitro target binding assay (selectivity against family members)

2o. Cell based blocking assay

3o. Cell based functional assay

Hit subcloning and characterization

Reagent preparation

Assay Development

Hybridoma library: 5-50x104 clones in 50-100 plates/ fusion, total 4-8 fusions

Animal Immunization

Lead selection

Animal model study (efficacy)

Lead optimization (Humanization, affinity maturation if necessary)

Target engagement, PK and safety assessment

Criteria of Successful Lead:

Neutralizing activity

Binding affinity of sub-nM

Selectivity > 100 folds

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Case Study: Anti-Immune Checkpoint Lead Generation

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Note: the marketed drug was used as positive control

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E. coli Expression Services Pricing

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BacPowerTM Guaranteed Bacterial Protein Expression

Customer Provides Tag options Amount (mg) Timeline(weeks)

Purityoptions

Endotoxin level options Pricing

Target protein sequence

Small tag (e.g., His) 3-64-6 ≥75%

≥85%≥90%

≤1.0EU/µg≤0.1EU/µg

≤0.01EU/µgStarting from

$2,200Large tag (e.g., GST) 5-10

No Tag 5-10 7-8

Why use above service?Zero-risk, best value packages that can be used for most target proteins under 100kDa. Hassle free service and lots of options to choose from – we would recommend you try this first in case of an E. coli project. Moreover, you can get an

instant quote online within seconds

Introduction

BacPowerTM Duplex – Small Scale E. coli Protein Expression

Customer Provides Deliverables Purity Options Delivery Time Pricing

Target protein sequence• 3mg protein, guaranteed• COA including QC data• Gene sequence report

≥85% 4 weeks Under $1500/protein

When use above service?Ideal for proteins <500 aa without much customization *Not exhaustive list

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Service Amount Timeline Price Deliverable Notes

HTP Antibody Production

50µg6-8 weeks

$499/Ab • 1-step affinity purified Ab• Report & QC Data

Applicable to human IgG1, IgG4 and mouse

IgG1 κ light chains1mg $799/Ab

MamPowerTM

Guaranteed rAb 5mg-500mg 5-10 weeks $2999 to $20,000

• LC and HC in pUC57• Gene sequence report, QC

data• >95% Ab by SDS-PAGE or

HPLC• Endotoxin control*• Defined concentration*

Applicable to human IgG1, IgG4 and mouse IgG and their chimeric

Abs

HD Transient Expression

Expressiontest, 40ml

test, 100ml-1L protein

exp. and pur.

4-10 weeks $950 to $6900

• Expression evaluationpurified material depending upon chosen step

Comparison between regular and HD

platforms available

FragPowerTM Ab Fragments

10mg 7 weeks $2399 • Antibody fragment• Optimized gene sequence

report• QC data

For scFv and VHH/VHproduction in E.coli

3mg 10 weeks $3399 For Fab production in insect cells

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rAb Production Pricing Information

*Depends on service specifications#Not exhaustive list

Documents

GenScript Your Biology Research Partner · 2019. 6. 11. · GenScript, A Global Biology CRO. Global headquarters Piscataway, New Jersey, USA. Main production site Local technical