Viral Vector Generation

Cyagen+Viral Vector Generation

 

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For chimeric antigen receptor (CAR)-T cell therapy, the CAR molecule is the key to the effectiveness of the treatment. CAR molecules usually include extracellular structures such as single-chain variable regions and hinge regions of antibodies that recognize tumor antigens, transmembrane regions, and intracellular structures such as costimulatory signals and CD3ζ signal transduction regions. The design of CAR will influence the killing effect, in vivo persistence, and cytotoxicity of CAR-T cells. Therefore, for different tumor treatment targets, researchers need to design reasonable and effective CAR molecules and package them into CAR viruses for the subsequent construction of CAR-T cells.

 
Based on our rich experience in virus packaging platforms, Cyagen provides lentivirus (LV) packaging services with different quality standards, which are widely used in the construction of various cell line models, especially for difficult-to-infect T cells used in CAR-T cell construction. For CAR-T cell therapy, we also provide CAR molecular design services for different targets to meet the individual needs of CAR-T cell therapy researchers.
 

Our Viral Vector Generation Service

We provide a variety of CAR molecular design services, including second-generation, third-generation, and fourth-generation CAR design services, as well as personalized CAR molecular design services based on different cellular immunotherapies.

We also provide packaging services.

 

Type

Cell number

Titer

Turnaround

Lentivirus (LV)

 

CAR Lentivirus

≥×108 TU

≥1×108 TU/mL

   5-8 weeks

Conventional overexpression lentivirus

≥×108 TU

≥1×108 TU/mL

Conventional  interference lentivirus

≥×108 TU

≥1×108 TU/mL

Conventional knockout lentivirus

≥×108 TU

≥1×108 TU/mL

Purified Lentivirus

≥×109 TU

≥1×109 TU/mL

 

Advantages

  • Design of CAR molecules and vectors for various targets to meet individual needs.
  • A variety of vector catalogs shorten the experimental turnaround.
  • Mature lentivirus packaging technology, with many articles published by customers.
  • Strict virus quality control standards, high infection efficiency.
  • The third-generation lentivirus packaging system further improves biological safety.

 

Case 1. Lentiviral Titer via Flow Cytometry - CD19 Antigen

The lentiviral purification solution expressing CD19 antigen was diluted in four gradients (0.01, 0.1, 1, 10 μL) and added to the same number of 293T cells separately. After 72 hours, the positive rate of 293T cells was detected by flow cytometry. The percentage of virus-infected 293T cells to the total number of cells), and then calculate the transduction titer of the lentivirus according to the following formula:

A: The percentage of positive cells in the 0.1 μl group.   B: The percentage of positive cells in the 0.01 μl group.   N: Number of cells when virus is added   V: Volume of virus added in group A

 

图1. CD19抗原慢病毒滴度检测结果

The number of CD19 antigen-positive cells gradually increases with the increase of the transfection virus gradient, indicating that the lentivirus has infectious activity. The calculated virus titer is 1.4×108 TU/mL.

 

Case 2. Lentiviral Titer via Flow Cytometry - FMC63 CAR

The FMC63 CAR lentivirus titer was detected by the same method as in Case 1. After calculation, the virus titer is about 4.33×108 TU/mL.


图2. FMC63 CAR慢病毒滴度检测结果
 

Case 3. Images of Lentivirus-Infected Cells

HepG2
A549
HT1080
THP-1
Ploy
HepG2

Figure 3. The effects of lentivirus infection on various cells

Inquiries and Quote Requests

Request a quote now. Alternatively, you can always email animal-service@cyagen.com or call 800-921-8930 to inquire about our services or obtain a quote for your project.

 

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