Considerations in the development and validation of real-time quantitative polymerase chain reaction and its application in regulated bioanalysis to characterize the cellular kinetics of CAR-T products in clinical studies

被引：10

作者：

Yang, Tong-yuan ^{[1
]}

Doddareddy, Rajitha ^{[1
]}

机构：

[1] Janssen Res & Dev LLC, Janssen BioTherapeut, 1400 McKean Rd, Spring House, PA 19477 USA

来源：

BIOANALYSIS | 2021年 / 13卷 / 02期

关键词：

assay development and validation; CAR-T; cellular kinetics; qPCR; regulated bioanalysis; LIGAND-BINDING ASSAYS; PCR ASSAYS; CELLS; QUANTIFICATION; REVOLUTION; SUPPORT; DNA;

D O I：

10.4155/bio-2020-0221

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Real-time quantitative polymerase chain reaction (qPCR) has become the standard method for monitoring cellular kinetics of CAR-T therapies with measurement of the CAR transgene copy numbers in peripheral blood mononuclear cells isolated from patients receiving the treatment. Unlike other biophysical and immunological methodologies for bioanalytical characterization of conventional small molecule drugs or protein biologics, there is no relevant regulatory guidance to date on the method development and validation for quantitative qPCR assays employed during clinical development of CAR-T products. This paper will provide an overview and considerations in the development and validation of a qPCR assay from sample extraction to assay parameters and its implementation in regulated bioanalysis for CAR-T or other types of cell therapies.

引用

页码：115 / 128

页数：14

共 27 条

[11]

Geist BJ, 2013, BIOANALYSIS, V5, P227, DOI [10.4155/BIO.12.304, 10.4155/bio.12.304]

[12] Human Copy Number Variation and Complex Genetic Disease [J].

Girirajan, Santhosh ;

Campbell, Catarina D. ;

Eichler, Evan E. .

ANNUAL REVIEW OF GENETICS, VOL 45, 2011, 45 :203-226

[13]

Hedman Johannes, 2013, Methods Mol Biol, V943, P17, DOI 10.1007/978-1-60327-353-4_2

[14] Comparison of UV spectrometry and fluorometry-based methods for quantification of cell-free DNA in red cell components [J].

Khetan, Dheeraj ;

Gupta, Nitesh ;

Chaudhary, Rajendra ;

Shukla, Jai Shankar .

ASIAN JOURNAL OF TRANSFUSION SCIENCE, 2019, 13 (02) :95-99

[15] Challenges and opportunities in bioanalytical support for gene therapy medicinal product development [J].

Ma, Mark ;

Balasubramanian, Nanda ;

Dodge, Robert ;

Zhang, Yan .

BIOANALYSIS, 2017, 9 (18) :1423-1430

[16] Cellular kinetics of CTL019 in relapsed/refractory B-cell acute lymphoblastic leukemia and chronic lymphocytic leukemia [J].

Mueller, Karen Thudium ;

Maude, Shannon L. ;

Porter, David L. ;

Frey, Noelle ;

Wood, Patricia ;

Han, Xia ;

Waldron, Edward ;

Chakraborty, Abhijit ;

Awasthi, Rakesh ;

Levine, Bruce L. ;

Melenhorst, J. Joseph ;

Grupp, Stephan A. ;

June, Carl H. ;

Lacey, Simon F. .

BLOOD, 2017, 130 (21) :2317-2325

[17]

O'Hara DM, 2013, BIOANALYSIS, V5, P2679, DOI [10.4155/BIO.13.241, 10.4155/bio.13.241]

[18] CD19-targeted CAR T-cell therapeutics for hematologic malignancies: interpreting clinical outcomes to date [J].

Park, Jae H. ;

Geyer, Mark B. ;

Brentjens, Renier J. .

BLOOD, 2016, 127 (26) :3312-3320

[19] The Cellular Immunotherapy Revolution: Arming the Immune System for Precision Therapy [J].

Paucek, Richard D. ;

Baltimore, David ;

Li, Guideng .

TRENDS IN IMMUNOLOGY, 2019, 40 (04) :292-309

[20] ENZYMATIC AMPLIFICATION OF BETA-GLOBIN GENOMIC SEQUENCES AND RESTRICTION SITE ANALYSIS FOR DIAGNOSIS OF SICKLE-CELL ANEMIA [J].

SAIKI, RK ;

SCHARF, S ;

FALOONA, F ;

MULLIS, KB ;

HORN, GT ;

ERLICH, HA ;

ARNHEIM, N .

SCIENCE, 1985, 230 (4732) :1350-1354

← 1 2 3 →