Generating NK cell receptor-Fc chimera proteins from 293T cells and considerations of appropriate glycosylation.

Alon Zilka; Michal Mendelson; Benyamin Rosental; Oren Hershkovitz; Angel Porgador

doi:10.1007/978-1-60761-362-6_18

Generating NK cell receptor-Fc chimera proteins from 293T cells and considerations of appropriate glycosylation.

Alon Zilka, Michal Mendelson, Benyamin Rosental, Oren Hershkovitz, Angel Porgador

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2 Scopus citations

Abstract

The use of recombinant receptors as a scientific tool has become widespread in many research fields. Of particular interest are the natural killer (NK) receptors that play a major role in the immune response against tumors and virus-infected cells. We present here (i) a detailed protocol for the production and purification of soluble recombinant NK cell receptors tagged with human IgG1-Fc (thus termed receptor-Fc chimera or receptor-Ig fusion protein) and (ii) a protocol for cell staining with these recombinant receptor-Fc chimeras. As these recombinant proteins are produced in eukaryotic cells, we further discuss the glycosylation pattern of these receptors that might interfere with their ligand-binding phenotype.

Original language	English
Pages (from-to)	275-283
Number of pages	9
Journal	Methods in molecular biology (Clifton, N.J.)
Volume	612
DOIs	https://doi.org/10.1007/978-1-60761-362-6_18
State	Published - 1 Jan 2010

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Molecular Biology
Genetics

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10.1007/978-1-60761-362-6_18

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AU - Zilka, Alon

AU - Mendelson, Michal

AU - Rosental, Benyamin

AU - Hershkovitz, Oren

AU - Porgador, Angel

PY - 2010/1/1

Y1 - 2010/1/1

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AB - The use of recombinant receptors as a scientific tool has become widespread in many research fields. Of particular interest are the natural killer (NK) receptors that play a major role in the immune response against tumors and virus-infected cells. We present here (i) a detailed protocol for the production and purification of soluble recombinant NK cell receptors tagged with human IgG1-Fc (thus termed receptor-Fc chimera or receptor-Ig fusion protein) and (ii) a protocol for cell staining with these recombinant receptor-Fc chimeras. As these recombinant proteins are produced in eukaryotic cells, we further discuss the glycosylation pattern of these receptors that might interfere with their ligand-binding phenotype.

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JO - Methods in molecular biology (Clifton, N.J.)

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Generating NK cell receptor-Fc chimera proteins from 293T cells and considerations of appropriate glycosylation.

Abstract

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