Antigen density quantification of cell-surface immunotherapy targets by flow cytometry: Multi-antigen assay of neuroblastoma bone marrow metastasis

Molly T Radosevich; Scott J Bornheimer; Majid Mehrpouryan; Bita Sahaf; Jean S Oak; Crystal L Mackall; Sabine Heitzeneder

doi:10.1016/j.xpro.2023.102709

Antigen density quantification of cell-surface immunotherapy targets by flow cytometry: Multi-antigen assay of neuroblastoma bone marrow metastasis

STAR Protoc. 2023 Dec 15;4(4):102709. doi: 10.1016/j.xpro.2023.102709. Epub 2023 Nov 17.

Authors

Molly T Radosevich¹, Scott J Bornheimer², Majid Mehrpouryan², Bita Sahaf¹, Jean S Oak³, Crystal L Mackall⁴, Sabine Heitzeneder⁵

Affiliations

¹ Center for Cancer Cell Therapy, Stanford Cancer Institute, Stanford University School of Medicine, Lorry Lokey Building, Suite G3141, MC: 5456, 265 Campus Drive, Stanford, CA 94305, USA.
² BD Biosciences, San Jose, CA 95131, USA.
³ Department of Pathology, Stanford University School of Medicine, Stanford, CA, USA.
⁴ Center for Cancer Cell Therapy, Stanford Cancer Institute, Stanford University School of Medicine, Lorry Lokey Building, Suite G3141, MC: 5456, 265 Campus Drive, Stanford, CA 94305, USA; Department of Pediatrics, Stanford University School of Medicine, Stanford, CA 94305, USA; Parker Institute for Cancer Immunotherapy, San Francisco, CA 941209, USA; Department of Medicine, Stanford University School of Medicine, Stanford, CA 94305, USA.
⁵ Center for Cancer Cell Therapy, Stanford Cancer Institute, Stanford University School of Medicine, Lorry Lokey Building, Suite G3141, MC: 5456, 265 Campus Drive, Stanford, CA 94305, USA; Department of Pediatrics, Stanford University School of Medicine, Stanford, CA 94305, USA.

Abstract

The central role of target antigen density on chimeric antigen receptor T cell potency highlights the need for accurate measurement of antigen levels on clinical tumor samples. Here, we present a protocol for quantifying antigen density for six cell-surface antigens on neuroblastoma cells metastatic to bone marrow. We describe steps for patient sample acquisition, flow cytometry panel development, instrument setup, and compensation and detail procedures for running clinical samples and data analysis. For complete details on the use and execution of this protocol, please refer to Heitzeneder et al. (2022).¹.

Keywords: Cancer; Flow Cytometry; Immunology.

MeSH terms

Bone Marrow
Bone Marrow Neoplasms* / therapy
Flow Cytometry
Humans
Immunotherapy
Neuroblastoma* / therapy

Abstract

MeSH terms

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