PerkinElmer
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Application Note

A Comparative Study of Two Immunoassay Platforms to Determine Lentivirus Titer for CAR-T Development

Introduction

The main vectors of gene therapy in research are viruses. The most popular tool for gene delivery is a genetically modified lentivirus. Modified lentivirus (HIV-1) vectors retain their ability to infect undivided cells, thereby increasing their ability to transduce a wide variety of cells, including those that are difficult to transduce. This advantage enables the stable long-term expression of a transgene.

In immunotherapy, CAR-T cells are manufactured by transducing the CAR gene with an HIV-1 vector in T cells to express a specific chimeric p24 protein on their surface. This allows them to recognize cancer cells and destroy them. These CAR-T cells must be generated individually to treat each patient.

This application note demonstrates a comparative quantification of the p24 titer in a lentiviral GFP control sample using Alliance HIV-1 p24 Antigen ELISA and p24 AlphaLISA immunoassay platforms.

Check out the different sections of this application note:

  • A lentiviral vector that encodes the CAR construct
  • Determination of the efficiency of transient co-transfection by measuring viral titer
  • Detection of the presence of a p24 HIV-1 specific antibody in the sample
  • Quantification of targets present in the sample