Your Partner in Biologics Discovery� and Development

www.lakepharma.com

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About LakePharma

• Dedicated client service and project management team

• 24/7 secure access to cloud-based client portal

• 100% CRO business model, no internal drug development programs

• Founded in 2009• Acquired Blue Sky BioServices in March, 2016• All services performed in the US (San Francisco,

CA; Cambridge/Boston, MA)• Key technologies developed in-house,

state-of-the-art facilities and instruments

LakePharma is a leading contract research organization (CRO) specializing in antibody and protein engineering, production, and cell line development.

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• Gene synthesis – all constructs fully sequence-verified• Molecular cloning:

- Ligation, PCR, site mutagenesis and other custom cloning

• Stand-alone DNA sequencing analysis• Plasmid DNA production:

- Midi, Maxi, Mega, Giga - High-throughput 96-well miniprep - Low endotoxin service

• 23-hour DNA purification• Gene expression and copy number analysis

LakePharma offers all recombinant DNA services, from gene synthesis to DNA purification.

Molecular Biology�

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• UberCell™ immunizations for functional mAbs against membrane targets

• DNA based or rapid protein based RIMMS immunizations

• Tolerance-breaking immunizations• Trianni humanized mouse platform – selecting

human antibodies in vivo• Electrofusion for high fusion efficiency• High-throughput 384-well screening:

- Incorporation of functional assays in screening cascade

• Delivery of clonal, functional mAbs with V-region sequences, hybridomas, and functional data

Therapeutic grade monoclonal antibody development in vivo

Antibody� Discovery�

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• Generation of phage libraries from immunized animals or responders

• SdAb, ScFv or Fab phage library construction and screening

• Single B-cell cloning• Hybridoma cloning and sequencing• Hybridoma scale-up and production• IgG reformatting to encode different

constant regions

Alternative approaches and additional services to generate novel monoclonal antibodies

Antibody� Discovery�

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• Customized library construction and screening• Epitope binning and mapping• Antibody humanization• Affinity maturation • Affinity measurement• Fcγ receptor panel

and FcRn binding assay• Therapeutic

developability analysis

LakePharma’s Antibody Engineering team provides end-to-end services, from design, production, to functional screening.

Antibody� Engineering

Antibody� Production

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LakePharma has extensive experience with constructing and producing antibodies in our mammalian HEK293 and CHO platforms. • Antibody reformatting to encode different

constant regions• Proprietary high-expression vectors• Transient and stable expression

in LakePharma-developed HEK293 and CHO cell lines

• Flexible scales, yielding mg- to gram-level antibodies

• High-throughput 96-well block production• IgG, bi-specific antibody, Fab, ScFv, IgM,

and more

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• Optimized titer-less expression with IKM saves weeks in expression testing

• Titer-less Infected Cell Preservation and Scale-up (TIPS) provides a better way to preserve and scale up baculovirus

• Baculovirus Infected Insect Cell (BIIC) offers superior stability and quality of infection

• Expression scouting to optimize quality and yield

• Variable scales (10mL to 300L) with industry leading turnaround times

• Experienced team led by William Hermans who pioneered these technologies

Insect expression: proprietary infection kinetics monitoring (IKM) provides unmatched scalability and reproducibility.

Protein Production

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• Transient and stable transfection in suspension HEK293 or CHO cell lines

• Flexible production volumes and custom purification methods

• Proprietary high-expression vectors for optimized protein production system

• E. coli strain creation and expression optimization

• Scale up services up to 100 L in flask or fermentor

• Strain creation and expression optimization• Scale up services up to 100 L in flask or fermentor• Pichia pastoris, Saccharomyces cerevisiae

Mammalian expression:

Bacterial expression:

Yeast expression:

Protein Production

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• Protein cleavage• Biotinylation• Avi tag: in vitro and in vivo services• HRP conjugation• Fluorophore conjugation• Bead conjugation• Additional custom conjugation services

are available upon request

LakePharma offers custom conjugation and chemical modification services for proteins and antibodies.

Protein Chemistry�

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• Cell lines suited for in vitro assays: rapid and cost-efficient in a variety of host cell lines via non-viral and viral-based approaches

• CHO stable cell lines for bioproduction of proteins and antibodies: suitable for small to mid-scale manufacturing

• Biologics production cell lines to ensure the highest yield and quality: - Freedom to operate - Suspension-adapted CHO-GSN cells - Uses chemically defined media without serum or other animal-derived products - Suitable for cGMP manufacturing

LakePharma provides 3 types of stable cell line generation services.

Cell Line Development

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• Upstream and downstream optimization• Demonstrate clonal cell-line stability• Produce and test Master Cell Banks (MCB)• Develop a high-performance process suitable

for clinical supply: - Transfer process to CMO

• Develop formulation suitable for use in GLP tox and Phase I studies

• Develop and qualify analytical assays for stability and lot release

• Generate stability data to support shelf-life of CTM

• Characterize reference materials• Support IND documentation

Process Development

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• Formulation development: - SE-HPLC, DSC/DSF, cIEF, and other methods as required - Includes forced degradation study - Viscosity for high concentration formulation

• Stability study: - SE-HPLC, cIEF, CE-SDS, binding/bioassay and other methods

• Comparability analysis: - Intact mass spec, sequence confirmation, PTM analysis

• Glycosylation analysis• de novo antibody sequencing• Method development and qualification

LakePharma uses the latest advances in protein analytical services to provide protein characterization.

Bioanaly�tics

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• Primary and differentiated cell assays• High content screening• Small molecule cytotoxicity – HepG2• Cytokine stimulation and release• Flow cytometry and FACS

• ELISA• Luminescence• Fluorescence - FP, HTRF, FRET, TR-FRET• SPA and radioligand binding• LCMS-based analyses• qPCR - relative and absolute quantitation

Cell-based assays Assay formats/readouts

Fully� Customized Assay� Design and Development

Multiple Technology Platforms, Antigen Creation, Hybridoma Development, Hybridoma Cloning &

Screening, High-throughput Hybridoma Sequencing, Phage Library Construction & Screening,

and Single B-Cell Cloning

Antibody Discovery

Stable Cell Line Development, Master Cell Banking, Cell Culture

Development & Optimization, Custom Media Development,

Purification Process Development & Scalability,

and Production Scale-up

Cell Line & Process Development

Kinetic Measurements and Biophysical Characterization including Intact Mass (LC-MS), Sequence Confirmation,

Post Translational Modification, Protein Aggregation (SE-HPLC), Thermostability (DSC), Particle Size (DLS),

Charge Variation (IEX), pI, RP-HPLC, CE-SDS, and formulation & Stability Analysis

Bioanalytics

Plasmid Design, Gene Synthesis, Cloning, DNA Sequence Analysis, Plasmid Production, TeraPrep™ Transfection-Grade Plasmid Preps, and Viral Preps

Molecular Biology

Antibody Humanization, Affinity Maturation, de novo Antibody Sequencing, Proprietary High-expression Vectors, Expression Using Microbial, Mammalian, Insect & Yeast Systems, and Production of a Variety of Proteins, Antibodies, Antibody-like Molecules & Fusion Proteins

Antibody Engineering & Production

Biochemical and Cell-based Assay Development & Screening, Radioligand Binding, ELISAs, Meso Scale Discovery Assays, High Content Screening, Proliferation & Viability Assays, Flow Cytometry, and qPCR

Pharma Assay Development & Screening