Quantitative analysis by LBA (PK/Biomarker) 1
Seiji Mito (Mitsubishi Tanabe Pharma Corporation) Kousuke Iijima (Kyowa Hakko Kirin Co., Ltd.) Satomi, Sasahara (JCL Bioassay Corporation) Hiroyuki Shimizu (Toray Research Center, Inc.) Masako Soma (Daiichi Sankyo Co., Ltd.) Fumiaki Meguro (LSI Medience Corporation) Itadaki Yamaguchi (Sumika Chemical Analysis Service, Ltd.) Yoshinobu Yokota (Shin Nippon Biomedical Laboratories, Ltd.) 2
Biotechnology-based drugs, typified by antibody drugs, are currently being developed for various diseases. The ligand binding assay (LBA) is being used for the concentration measurement of these drugs. LBA is also commonly used to measure various biomarkers that serve as indices for drug efficacy and safety evaluations. In this regard, LBA was taken up as one of the topics in a JBF discussion group (DG) starting in 2013. Presentations were held in the fourth and fifth JBF symposium. As to this particular topic, the discussion was held with a focus on the quantitative analysis by LBA (PK/Biomarker). 3
In particular, this DG talked about sample preparations, LBA platforms, surrogate matrices, stability assessments, specificity, reference standards and critical reagents. This presentation gives an overview of contents discussed in the DG to help establish the standard procedures for the quantitative analysis by LBA (PK/Biomarker) and reliability of the assay. 4
May 2014: Member recruitment from DG supporters. 24 Jun 2014: Kick off meeting with 8 members. Jul 2014 - Dec 2014: Monthly teleconferences and e-mail conversations. 20 Aug 2014: F2F meeting collaborated with DG2014-11. Jan 2015: Summarizes and preparation poster presentation. 5
1. Sample preparation 2. LBA platform 3. Surrogate matrix 4. Stability evaluation 5. Specificity 6. Reference standard Critical reagent 6
1.1 Operating and inspecting pipette 1.2 Using low-protein binding tip and tube, and sample additives 7
Back ground Discussion points Operation Do you use both pipetting modes (forward or reverse mode)? How to select the pipetting mode and the volume range Inspection When to perform the test Test volume 8
Back ground Discussion points When to select a low-protein binding tip and tubes Do you use the same tip and tube in actual sample analysis? When to select to use sample additives Do you use the same additives in actual sample analysis? 9
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Background Discussion points Commonly used Platforms Priority criteria of Platform selection Risk management of using a Platform supported by a single vendor What should be done when a validated assay have to be changed its platform. Will you continue to use an instrument with expired warranty terms? 11
1.1 Selection of a surrogate matrix 1.2 Analytical method validation and stability 12
Back ground Discussion points What is used as a surrogate matrix? How to select the surrogate matrix Risk management of using a surrogate matrix 13
Back ground Discussion points Analytical method validation Stability 14
4-1) Considerations for Validation Study Stability Evaluations 4-2) Importance of ISS 4-3) Stability evaluation for Biosimilar 15
Background Discussion points 4-1) Considerations for Validation Study Stability Evaluations Target-relevant stability samples (PK or biomarkers) Stability conditions and period Stability evaluation method using individual sample Stability evaluation method using SS*-spiked sample Different criteria for PK and biomarkers Handling against incidental criterion non-fulfillment 4-2) Importance of ISS 4-3) Stability Evaluation for Biosimilars *: Standard substance 16
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Background Discussion points Related substances and selection When do you not perform the specificity? Setting of spiked concentration When a commercial kit is used, Is the specificity necessary? When a related substance exists in a matrix Judgment of necessity for additional specificity testing after a method validation is completed 18
6.1 Stability of purchased reference standard 6.2 Handling of lot changes 6.3 Expiration date of critical reagent 19
Background Discussion points Stability of purchased reference standard Handling of lot changes Reference standard Critical reagent Assay kit Expiration date of critical reagent 20
LBA (PK/Biomarker) 21
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ligand binding assay (LBA) LBA 2013 JBF (DG) LBA 4 5 JBF LBA PK/Biomarker 23
DG LBA DG LBA PK/Biomarker 24
2014 5 DG 2014 6 24 8 2014 7 2014 12 1 TC 2014 8 20 DG2014-11 F2F 2015 1 25
1. 2. LBA platform 3. 4. 5. 6. 26
1.1 1.2 27
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LBA 29
1 2 1 2 2 30
Japan Bioanalysis Forum QC 8 31
Japan Bioanalysis Forum QC QC QC QC 揃 32
Japan Bioanalysis Forum + + + 33
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Japan Bioanalysis Forum CMC TK PK PK 36
TK PK PK 37
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Japan Bioanalysis Forum Platform Platform Single vendor Platform 39
Japan Bioanalysis Forum LBA 40
Immuno PCR SPR TMB, pnpp Luminol, dioxetane Ruthenium Alexa Fluor, Europium DNA SPR Immuno PCR SPR 41
platform PK ELISA ECL ELISA ECL Biomarker ELISA 42
Single vendor LBA platform platform 43
platform 44
PK Biomarker 45
1.1 1.2 46
Japan Bioanalysis Forum 47
Japan Bioanalysis Forum 4.1. 48
Japan Bioanalysis Forum. * * * 49
Japan Bioanalysis Forum 50
Japan Bioanalysis Forum 51
Japan Bioanalysis Forum 52
Japan Bioanalysis Forum. 53
Japan Bioanalysis Forum. n 5. 54
Japan Bioanalysis Forum...... MRD.. 55
Japan Bioanalysis Forum. QC.. ISS Incurred Sample Stability. ISS ISR. 56
4-1) 4-2) ISS 4-3) 57
Japan Bioanalysis Forum 4-1) PK, Biomarker PK Biomarker 4-2) ISS 4-3) 58
Japan Bioanalysis Forum LBA PK * Biomarker fit-for-purpose** PK Biomarker *: ( 0711 1 ) **: Fit-for-Purpose Method Development and Validation for Successful Biomarker Measurement ( AAPS Research paper, 2005 ) 59
/ PK, Biomarker PK LQC* HQC** Extra QC n=3/ Biomarker PK LQC HQC CRO 5 7 PK Biomarker *: Low quality control, within 3 times of LLOQ (lower limit of qualification) **: High quality control, more than one-thirds of ULOQ (upper limit of qualification) 60
/ PK, Biomarker PK Biomarker 4 24 3 5 1 3 6 4 24 PK Biomarker Deep Freezer -80-20 -80 61
1 5-7 7 4 %Initial* 20% *: %Initial = / 100 62
PK LQC, HQC, Extra QC 20% PK Biomarker LQC, HQC, Extra QC LQC, HQC Extra QC / 20% LQC HQC Extra QC* 20% 20% 20% *: BMV Extra QC 8 4 50% 50% 63
PK Biomarker PK 20% Biomarker PK 20% Biomarker 20% 25% Biomarker 64
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24 4 5 3 LQC, HQC, Extra QC HQC n=3/ 3 LQC Extra QC HQC n=1 n=3 CV 20% 僅 僅 66
1 2 3 2 1,3,6 2 1,3,6 3 HQC 15% 6 3 2 6 15% 20% %Initial 20% 67
PK Biomarker ISS* PK ISR** ISS Biomarker ISS PK ISS *: Incurred sample stability **: Incurred sample reanalysis 68
PK 1 69
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Japan Bioanalysis Forum 71
Japan Bioanalysis Forum 72
IgG IFN IFN-α β 73
Japan Bioanalysis Forum a. b. COA 74
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PK 76
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6.1 6.2 6.3 80
Japan Bioanalysis Forum 81
Japan Bioanalysis Forum Q&A No.3 Q&A No.25 82
Japan Bioanalysis Forum 83
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/PK 85
/PK QC QC QC QC 5 (LLOQ L M H ULOQ). 3 (L M H) 86
PK BM PK BM PK 87
QC,, ( ) QC 5 (LLOQ L M H ULOQ) 3 (L M H). 88
/PK QC (5 LL L M H UL) 89
/ BM PK 90
Q&A No.25 QC 91
QC 10 92