Sample Assay Report. Immunotherapy Inhibitor Assays Sample Report. Page 1 of 13

Transcription

1 Sample Assay Report Immunotherapy Inhibitor Assays Sample Report Page 1 of 13

2 Immunotherapy Inhibitor Assays Study Sponsor: Attention: Address: Study Director: Testing Facility: Henry Zhu, Ph.D. BPS Bioscience Inc Cornerstone Court West, Ste. B San Diego, CA USA Study Period: Report Version: 1 Report Date: Page 2 of 13

3 Study Director Aaron Snead Senior Scientist II, Ph.D. Date Henry Zhu, Ph.D. President Date Page 3 of 13

4 CONTENTS IMMUNOTHERAPY INHIBITOR ASSAYS... 1 IMMUNOTHERAPY INHIBITOR ASSAYS... 2 STUDY DIRECTOR PURPOSE OF THE STUDY MATERIALS AND METHODS MATERIALS COMPOUNDS EXPERIMENTAL CONDITIONS Proteins and Substrates Assay Conditions Data Analysis ASSAY RESULTS SUMMARY OF THE INHIBITORY EFFECTS OF THE COMPOUNDS ON IMMUNOTHERAPY INHIBITORY ASSAYS RESULTS OF THE EFFECTS OF THE COMPOUNDS ON IMMUNOTHERAPY ASSAYS RESULTS OF THE EFFECTS OF THE COMPOUNDS ON IMMUNOTHERAPY ASSAYS TABLE STORAGE AND RETENTION OF RECORDS QUALITY ASSURAE STATEMENT Page 4 of 13

5 1. Purpose of the Study The purpose of the study is to determine the effect of test compounds from Sample Report on the activity of various immunotherapy targets. Page 5 of 13

6 2. Materials and Methods 2.1 Materials Protein Catalog # Lot # PD D PD-L1-biotin PD-1 mab D PD-L2-biotin PD-L D B7-1-biotin B PD-L1 mab D BTLA D HVEM-biotin BD HVEM CD CTLA CTLA4 Ab D CD SIRPα-biotin SIRPα OX40L D OX40-biotin OX Compounds Compound I.D. Compound Supplied Stock Concentration Dissolving Solvent Test Range Page 6 of 13

7 2.3 Experimental Conditions Proteins and Substrates Protein Catalog # Lot # Protein Used /reaction PD D 100 ng PD-L1-biotin ng PD-1 mab D 30 nm ref PD-L2-biotin ng PD-L D 250 ng B7-1-biotin B ng PD-L1 mab D 30 nm ref BTLA D 100 ng HVEM-biotin BD 100 ng HVEM μm ref CD ng CTLA ng coat (1 μm ref) CTLA4 Ab D 30 nm ref CD ng SIRPα-biotin ng SIRPα μm ref OX40L D 200 ng OX40-biotin ng OX nm ref Assay Conditions Experimental conditions where done according to the Respective Binding Assay Kit Protocol ( Coat proteins were added to the plate 50 L at 2-5 ng/ L at 4 ºC overnight. Test compounds or reference compound/neutralizing antibody were added to the coated plate followed by addition of the corresponding biotinylated binding partner. Reaction was incubated for 2 h at room temperature. Page 7 of 13

8 2.3.3 Data Analysis Binding assays were performed in duplicate at each concentration. The luminescence data were analyzed using the computer software, Graphpad Prism. Percent inhibition was determined by normalizing the data to signal from Negative Control wells (uncoated wells treated with the biotinylated ligand, set as 100% inhibition) and Positive Control wells (coated wells treated with the biotinylated ligand in the absence of any inhibitor, set as 0% inhibition). Data for a reference compounds or antibodies are included as a control for inhibition. Page 8 of 13

9 3. Assay Results 3.1. Summary of the Inhibitory Effects of the Compounds on Immunotherapy Inhibitory Assays Page 9 of 13

10 3.2. Results of the Effects of the Compounds on Immunotherapy Assays Results of the Effects of the Compounds on Immunotherapy Assays Table Activity of Compounds Assay Condition Concentration Raw Data % Activity Repeat 1 Repeat 2 Repeat 1 Repeat 2 Negative Control PD-1:PD-L1 Positive Control erence Inhibitor 30 nm PD-1 Ab Negative Control PD-1:PD-L2 Positive Control erence Inhibitor 30 nm PD-1 Ab Negative Control PD-L1:B7-1 Positive Control erence Inhibitor 30 nm PD-L1 Ab Negative Control BTLA:HVEM Positive Control erence Inhibitor 1 µm HVEM Negative Control CD28:B7-1 Positive Control erence Inhibitor 100 nm CTLA Negative Control CTLA4:B7-1 Positive Control erence Inhibitor 30 nm CTLA4 Ab Negative Control CD47:SIRPα Positive Control erence Inhibitor 30 µm SIRPα Negative Control OX40:OX40L Positive Control erence Inhibitor 5 µm OX Page 10 of 13

11 % Activity Sample Immunotherapy Binding Data PD-1:PD-L1 PD-1:PD-L2 PD-L1:B7-1 BTLA:HVEM CD28:B7-1 CTLA4:B7-1 CD47:SIRP OX40:OX40L Page 11 of 13

12 Storage and Retention of Records The original final report provided to the sponsor will be kept by the sponsor under its sole responsibility. Page 12 of 13

13 5. Quality Assurance Statement I certify that the results presented in this report were generated using the materials and methods mentioned and that these results reflect the Raw Data. Henry Zhu, Ph.D. President Date Page 13 of 13