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reverse phase high performance liquid chromatography hplc column  (Restek Corporation)

 
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    Restek Corporation reverse phase high performance liquid chromatography hplc column
    Reverse Phase High Performance Liquid Chromatography Hplc Column, supplied by Restek Corporation, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/reverse phase high performance liquid chromatography hplc column/product/Restek Corporation
    Average 86 stars, based on 1 article reviews
    reverse phase high performance liquid chromatography hplc column - by Bioz Stars, 2026-06
    86/100 stars

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    A Upper panel depicts the large-scale workflow used for the stable transfection and processing of B lymphoblastoid cell lines (BLCL) cells using EF1α-GoI-pIRES-DsRed plasmids containing SARS-CoV-2 genes. Cell pellets (4 x 10 8 to 1 x 10 9 ) were homogenised using a cryomill, lysed, and peptide-HLA complexes immunoaffinity purified using first the W6/32 antibody (HLA class I) and subsequently a mix of SPV-L3, LB3.1, B7/21 antibodies (HLA class II). Peptides were separated from HLA using reverse-phase high-performance liquid <t>chromatography</t> <t>(HPLC).</t> B The lower panel shows the workflow schematic for small-scale protein antigen direct delivery experiments. SARS-CoV-2-derived proteins were electroporated into BLCL and 0.9 ×108 to 1.6 × 108 cells collected 48 h later. Cell pellets were subjected to direct lysis and immunoaffinity purification. Peptides were separated from HLA using 5 kilodalton (kDa) molecular weight cut-off filters. All samples were analysed using liquid chromatography-tandem mass spectrometry (LC-MS/MS) and PASEF® on TimsTOF Pro. MHC: Major histocompatibility complex. Created with BioRender.com released under a Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International license.
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    A Upper panel depicts the large-scale workflow used for the stable transfection and processing of B lymphoblastoid cell lines (BLCL) cells using EF1α-GoI-pIRES-DsRed plasmids containing SARS-CoV-2 genes. Cell pellets (4 x 10 8 to 1 x 10 9 ) were homogenised using a cryomill, lysed, and peptide-HLA complexes immunoaffinity purified using first the W6/32 antibody (HLA class I) and subsequently a mix of SPV-L3, LB3.1, B7/21 antibodies (HLA class II). Peptides were separated from HLA using reverse-phase high-performance liquid chromatography (HPLC). B The lower panel shows the workflow schematic for small-scale protein antigen direct delivery experiments. SARS-CoV-2-derived proteins were electroporated into BLCL and 0.9 ×108 to 1.6 × 108 cells collected 48 h later. Cell pellets were subjected to direct lysis and immunoaffinity purification. Peptides were separated from HLA using 5 kilodalton (kDa) molecular weight cut-off filters. All samples were analysed using liquid chromatography-tandem mass spectrometry (LC-MS/MS) and PASEF® on TimsTOF Pro. MHC: Major histocompatibility complex. Created with BioRender.com released under a Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International license.

    Journal: Nature Communications

    Article Title: Mapping the immunopeptidome of seven SARS-CoV-2 antigens across common HLA haplotypes

    doi: 10.1038/s41467-024-51959-6

    Figure Lengend Snippet: A Upper panel depicts the large-scale workflow used for the stable transfection and processing of B lymphoblastoid cell lines (BLCL) cells using EF1α-GoI-pIRES-DsRed plasmids containing SARS-CoV-2 genes. Cell pellets (4 x 10 8 to 1 x 10 9 ) were homogenised using a cryomill, lysed, and peptide-HLA complexes immunoaffinity purified using first the W6/32 antibody (HLA class I) and subsequently a mix of SPV-L3, LB3.1, B7/21 antibodies (HLA class II). Peptides were separated from HLA using reverse-phase high-performance liquid chromatography (HPLC). B The lower panel shows the workflow schematic for small-scale protein antigen direct delivery experiments. SARS-CoV-2-derived proteins were electroporated into BLCL and 0.9 ×108 to 1.6 × 108 cells collected 48 h later. Cell pellets were subjected to direct lysis and immunoaffinity purification. Peptides were separated from HLA using 5 kilodalton (kDa) molecular weight cut-off filters. All samples were analysed using liquid chromatography-tandem mass spectrometry (LC-MS/MS) and PASEF® on TimsTOF Pro. MHC: Major histocompatibility complex. Created with BioRender.com released under a Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International license.

    Article Snippet: For large-scale elution, the peptide-HLA mixtures were fractionated off-line using a 4.6-mm × 100-mm monolithic reversed-phase C18 high-performance liquid chromatography (HPLC) column (Chromolith SpeedROD; Merck Millipore) and an ÄKTAmicro HPLC system (GE Healthcare).

    Techniques: Stable Transfection, Purification, High Performance Liquid Chromatography, Derivative Assay, Lysis, Immunoaffinity Purification, Molecular Weight, Liquid Chromatography, Mass Spectrometry, Liquid Chromatography with Mass Spectroscopy, Immunopeptidomics