CAR-T Cell Line Characterization

All products and services are For Research Use Only and CANNOT be used in the treatment or diagnosis of disease.

As a leading cell therapy biotechnology company, Creative Biolabs provides one-stop CAR-T therapy development services, ranging from GMP/GLP-graded CAR plasmid production (providing mg-level to g-level production scale), GMP-graded CAR lentivirus production, to CAR -T cell production to meet the needs of non-clinical research and clinical phase I trials initiated by cellular therapy researchers.

Understanding each step of the CAR-T workflow is essential to ensure the quality and effectiveness of the final CAR-T product. An Ideally panel for new CAR-T development and QC, the CAR-T generation cell line characterization panel can help simplify the CAR-T manufacturing workflow and potentially reduce vein to vein time. Besides, contaminant-free cell banks lay a solid foundation for you to maintain a stable production path in the next few years. Therefore, proper cell line characterization is critical to confirm the certainty of the contamination-free cell bank. Creative Biolabs offers a wide range of CAR-T cell line characterization services, including phenotypic or genotypic identity testing, which is required for mammalian, microbial, and insect cell substrate testing to generate CAR-T cells.

Overview of our Cell Line Characterization Package

The characterization of CAR-T cell lines mainly involves the following cell lines:

CAR plasmid manufacturing cell line. Generally, bacteria or fungal cell lines are used to prepare GMP-grade plasmids through large-scale fermentation.
CAR lentivirus manufacturing cell line. By transfecting CAR lentivirus to 293T cell line to prepare cGMP grade bulk virus.
CAR-T cell manufacturing cell line. Usually, human autologous T cells or Jurkat cell lines, which are stable cell lines developed by transducing CAR lentivirus.

More specifically, the characterization of CAR-T cell lines is to perform cell lines on the main cell bank (MCB), working cell bank (WCB) used for production. According to current regulations, the following aspects must be considered:

Origin and history of cell morphology and growth characteristics of cell lines
Cell identity
The purity of the cell line (ie, non-contaminated cells, microbial contamination, and adventitious virus contamination)
Tumorigenicity and carcinogenicity
Stability

When executing the assays, Creative Biolabs follows the guidelines recommend and using testing procedures that are validated to show their suitability for intended CAR-T purpose.

Advanced Analysis Modules Available for CAR-T Cell Line Characterization

The following table lists the recommended test panels for characterizing general mammalian and microbial cell lines used to produce CAR-T, as well as T cells.

Microbial Cell Line Characterization Panel

Recommended Testing Items	MCB	WCB
Bacteriophage detection	+	+
Purity	+	+
Identification	+	+
Viability	+	+
Copy number	+	+
DNA Sequencing	+	+
Retention of selectable markers	+	+
Retention of recombinant construct	+	+
Restriction Endonuclease Analysis	+	+

Mammalian Cell line Characterization Panel

Recommended Testing Items		MCB	WCB	EPC/CAL
Microbial Contamination	Sterility	+	+	+
	Mycoplasma	+	+	+
	Mycobacterium	+
	Spiroplasma	+
Cell Line Identity	DNA Barcoding	+	+	+
	DNA fingerprinting/STR	+		+
	Karyology	+		+
Genetic stability	DNA sequencing	+		Optional
Genetic stability	Gene copy number by qPCR/STR	+		+
Virus Testing	Retroviruses: Reverse transcriptase and retroviral infectivity assays	+		+
	In vitro adventitious viruses	+	+	+
	In vivo adventitious viruses	+		+
	Test for specific viruses such as: Bovine viruses Porcine viruses Human viruses Minute virus of mice	+
	In vivo biosafety: Mouse, hamster, and rat antibody production assays (MAP, RAP, and HAP)	+
	Transmission electron microscopy	+		+
Tumorigenicity/Oncogenicity				+

T Cell line Characterization Panel

Recommended Testing Items		MCB	WCB	EPC/CAL
Microbial Contamination	Sterility	+	+	+
Microbial Contamination	Mycoplasma	+	+	+
SV40 Large T Antigen Sequence Analysis		+	+	+
Cell Line Identity	DNA Barcoding	+	+	+
Genetic stability	DNA sequencing	+		+
	Gene copy number	+		+
	Restriction endonuclease analysis	+		+
Virus Testing	Endogenous and non-endogenous retroviruses: reverse transcriptase and retroviral infectivity assays	+		+
	In vitro adventitious viruses	+		+
	In vivo adventitious viruses (suckling and adult mice, embryonated eggs)	+		+
	Test for specific viruses such as: Bovine viruses Porcine viruses Minute virus of mice	+
	In vivo biosafety: Mouse and hamster antibody production assays (MAP and HAP)	+
	Transmission electron microscopy	+		+