GenScript Your Biology Research Partner · 2019. 6. 11. · GenScript, A Global Biology CRO. Global...
Transcript of GenScript Your Biology Research Partner · 2019. 6. 11. · GenScript, A Global Biology CRO. Global...
GenScriptYour Biology Research Partner
September 28, 2017
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Company IntroductionMolecular Biology ServicesProtein ServicesAntibody ServicesPeptide ServicesADD Services
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Intellectual Property Protection
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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
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CRO Leadership
Quality Awards Innovation Awards Productivity Awards Reliability Awards
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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.
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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
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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
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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
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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
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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
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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
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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
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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
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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
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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.
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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
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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.
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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.
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Services Overview
GenScriptProducts & Services
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Gene
Peptide
Protein
Antibody
Discovery Biology
Cell Line
Thank youExpertise, Flexibility, Solutions
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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.
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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
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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
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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
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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