Verfahren zur Räumlichen Analyse von Genetisch Modifizierten Zelltherapieprodukten in Situ

Details

Veröffentlichungs-Nr.

EP4737896

Anmeldetag

4. November 2024

Veröffentlichung

6. Mai 2026

Rechtsraum

EP

IPC

G01N33/53, C12Q1/6841

Abstract

The invention is directed to a method to detect the phenotype and the spatial location of cells provided with a chimeric antigen receptor (CAR) in a fixed biological sample by targeting the mRNA coding for the chimeric antigen receptor (CAR) comprising the stepsa. providing a plurality of oligonucleotides, each having at least one detection section and at least one amplification section and two target binding sections at the respective 5' and 3' endsb. hybridizing the oligonucleotides with their 3' and 5' ends to the mRNAc. ligating the 3' and 5' ends of the hybridized oligonucleotides to generate circularized moleculesd. hybridizing an amplification oligonucleotide to at least one amplification section of the circularized molecules thus enabling rolling circle amplification (RCA) of the circularized moleculese. multiplying the circularized molecules by RCA in the fixed biological sample thereby creating DNA nanoballs comprising concatemers of the DNA nanoballsf. hybridizing detector oligonucleotides comprising at least one fluorescent unit to at least one detection section of the DNA nanoballs, thereby obtaining the spatial location of the cells provided with the chimeric antigen receptor (CAR) in the fixed biological sampleg. providing a plurality of fluorescently-labeled antibodies capable of binding to different phenotype defining antigens, thereby obtaining the phenotypes of the cells in the fixed biological sampleh. aligning the spatial location of cells provided with the chimeric antigen receptor (CAR) and the phenotypes of cells in the fixed biological sample.

Anmelder

Firma

Miltenyi Biotec B.V. & Co. KG

Land

🇩🇪 Deutschland

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