Exceptional Human Antibody Discovery. Corporate Overview

Transcription

1 Exceptional Human Antibody Discovery Corporate Overview Co 1 1

2 Our Mission Trianni is a biotech company with the scientific mission of creating optimized and highly versatile platforms for generation of fully-human monoclonal antibodies. 2

3 The Trianni Mouse TM Difference 3

4 The Trianni Mouse TM V H D H J H Cµ Mouse Locus Human V H Array D H J H Cµ 4

5 Mouse Machinery Drives Human Antibody Repertoire Production Promoter RSS Human V H gene segment Mouse genomic scaffold 5

6 TRN1/2/5 Core Alleles Heavy (H) 44 x V H D H J H Eμ μ δ γ3 γ1 γ2b γ2c ε α Kappa (K) 80% of Naïve Repertoire 39 x V K J K C K Lambda (L) Human 38 x V L J1C L J2C L J6C L J7C L J6C L J3C L Mouse 20% of Naïve Repertoire 6

7 Distinctive Features of Trianni s Technology Elimination of endogenous V, D and J gene segments Humanization of all three antibody loci H, K, L Designed immunoglobulin alleles Targeted insertion of arrays of synthetic antibody gene segments Optimizations throughout the arrays Chimeric gene segments (important re FTO) Mouse noncoding (regulatory) DNA Human coding DNA Platform flexibility Refinements can be generated quickly Alternative repertoire mice in development 7

8 In-House Platform Validation Antibody repertoire analysis All gene segments are used; CDR3 length and amino acid composition as in humans Immunization Ten model antigens tested (human extracellular protein domains) mabs similar to wild-type mice in terms of extent of somatic hypermutation, affinity, specificity and epitope coverage 8

9 Trianni Mice: Replete With B Cells and Antibodies WT B6 HHKK HHKKλλ CD19 TCRβ 9

10 Flow Cytometry of Spleen Cells from TRN H/k/l B220 Gated on: Total lymphocytes Igλ MZ B (CD21 Hi CD23 Lo ) CD19 Igκ CD23 Total B cells (CD19 + B220 + ) Igλ Fo B (CD21 + CD23 + ) 10 CD21 Igκ In gray, non-b cells in the same staining tube to illustrate the light chain staining backgrounds 10

11 A Highly Diverse Heavy Chain Repertoire V Gene IGHV3-13 IGHV4-28 IGHV3-53 IGHV3-20 IGHV3-48 IGHV4-34 IGHV3-74 IGHV1-8 IGHV1-69 IGHV3-30 IGHV3-38 IGHV1-2 IGHV4-4 IGHV1-18 IGHV3-11 IGHV1-46 IGHV4-39 IGHV3-33 IGHV4-61 IGHV3-23 IGHV1-24 IGHV V Gene IGHV1-3 IGHV3-72 IGHV1-58 IGHV2-70 IGHV3-9 IGHV5-51 IGHV3-73 IGHV3-16 IGHV3-35 IGHV3-43 IGHV1-45 IGHV3-66 IGHV3-49 IGHV3-15 IGHV2-5 IGHV4-31 IGHV2-26 IGHV7-81 IGHV3-64 IGHV3-7 IGHV6-1 IGHV Frequency (%) 44/44 VHs used Frequency (%) 11

12 H CDR3 Length Distribution 10 IgM-Sorted Naïve B cells Frequency (%) Mean: 15.3 Min: 0 Max: CDRH3 Length 12

13 Trianni Mouse CDR-H3 Composition Nearly Identical to Human 13

14 Evaluation, Large Pharma #1 Target 2, two campaigns (v1 and v2): both WT and Trianni Mice mount strong and similar immune responses 14

15 Anti-Target 2 Trianni Mouse mabs Demonstrate Superior Maximal and Average Potencies Compared to Wild-Type C57BL/6 Benchmark mabs 15

16 Evaluation, Large Pharma #2 - Two human cell surface proteins (Ag1 and Ag2) - A soluble human protein (Ag3) - Immunization: - Twice a week using Ribi adjuvant - A total of eight injections - Comparison to BALB/c mice - Three Phase Evaluation: - Immunization data - Immune response quality and diversity; fusion screening data - Detailed characterization of 20 mabs per target 16

17 Comparable Titers for All Three Antigens Trianni BALB/c 17

18 MAb Isolation Hybridoma generation Electrofusion of lymph node cells Primary screen Binding to human antigen Secondary screen Cross-reactivity to cynomolgus antigen Ligand-blocking activity Off-rates for all binders (using supernatants) Epitope competition (using supernatants) 18

19 Primary Screening Results Fusion parameters and primary binding similar to BALB/c 19

20 Secondary Screening Results Extended characterization of hu-ag+ supernatants similar to BALB/c Ligand blocking Cyno cross-reactivity Epitope coverage (Ag3 6/7 Trianni, 5/7 BALB/c) 20

21 Tertiary Analysis Subclone 20 hybridomas from each Tg animal/ag Determine productivity of hybridomas Characterize EC 50, IC 50 and K D for purified mabs Assess diversity of selected mabs 21

22 Data Summary Good quality antibodies were produced relative to control mice and benchmark antibodies Reasonable diversity seen with sequenced antibodies 22

23 TRIANNI Strongly Preferred in Side-by-Side Comparison Pharma #2 performed a side-by-side comparison with competitor human transgenic platform The Trianni Mouse performed much better the results were not even close. (Quote From Pharma #2) 23

24 Evaluation, Large Pharma #3 - Independent evaluation at two sites (US and Europe) - Multiple human antigens in both cases - Immunization: - RIMMS vs. conventional - Hybridomas and single B cell cloning - Antibody reformatting 24

25 Trianni Mouse-Derived Hybridoma Deliver Normal Production Yields and mab Quality 25

26 Purified Trianni MAbs Exhibit Sub-nM to nm Binding Affinities Affinity on Soluble Protein K d app (M) Affinity in subnm to nm range for both Trianni and BALB/c Mean ± SD (M) 4.8 x ± 5.4 x Mean ± SD (M) 3.9 x ± 9.2 x

27 Trianni mabs Are Routinely Reformatted into Fully-Human IgG Anti CD38 Clones Purified Batch Productivity mg/l Purity SDS-PAGE Mass spectrometry MW (Da) ELISA on CD38 EC50 (nm) LC HC Reformatted Hybridoma 109HHKK-3 VA % HHKK-3 VA > 99 % HHKK-3 VA % HHKK-3 VA > 99 % HHKK-3 VA % HHKK-5 VA % HHKK-5 VA % HHKK-7 VA % from reformatted Balb/c Sequence retrieved by RT-PCR from hybridoma Cloning VH and VL sequence into mammalian expression vectors Similar production yield and mab quality from reformatted mabs and Balb/c 27

28 Trianni mabs Display High Human Germinality Index Analysis from 33 paired VH/VL sequences, 10 VH germlines Germinality index (%) to human VH germlines

29 RIMMS Protocol Final Serum Titers 29

30 12 Week Rest-Boost Protocol Hybridoma Supernatants k off Ranking (Biacore) 30

31 In the Pipeline Mice that produce heavy-chain-only antibodies Mice with extended D regions that produce long CDR3-H3 sequences Mice in which plasma cells display immunoglobulin on the cell surface Mice that lack central B cell tolerance Mice that produce bispecific antibodies in vivo 31

32 Our Partners Below are the publically announced licensees of our 24 total partnerships DISCOVERY PARTNERS Janssen Magenta Mass General UAB Stanford 32

33 Why work with us? Small company means: - easy access to our experts - lean processes (quicker time to use of our platform) - personal relationships 60 years combined industry experience Representation in key biotech locations - San Francisco Bay Area, CA - Research Triangle Park, NC - Vienna, Austria Flexibility to create a custom partnership to match your needs Forward-thinking leadership Faster turn-around time Technical support, if needed An investment in your company. We value our partners and want you to succeed! 33

34 34 Contact Us