Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
6117095 | Immunology Letters | 2015 | 7 Pages |
Abstract
To test this assumption we used mRNA to express soluble derivatives of HLA-A2 in the human AF10 B cell myeloma and 624mel melanoma and H-2Kd in the mouse SP2/0 B cell myeloma. The level of MHC-I complexes secreted by these cells peaked within less than 24Â h post-transfection and they could be affinity-purified directly from the culture medium in considerably greater yields when compared to nonionic detergent lysates on a cell-to-cell basis. Mass-spectrometry analysis of eluted peptides revealed larger pools in the secreted material than in lysates with substantial overlap in composition. Our results introduce mRNA transfection as a powerful tool for determining the cell's MHC-I peptidome, which can be potentially applied to a broad range of cell types.
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Authors
Daphna Lazarus, Hadas Weinstein-Marom, Sigal Fishman, Rami Yossef, Dotan Zuri, Eilon Barnea, Arie Admon, Alon Margalit, Gideon Gross,