Cell membrane lipid with surface displaying GPI-NY-ESO-1 fusion protein and application thereof
A technology of GPI-NY-ESO-1 and NY-ESO-1, which is applied in non-active ingredients medical preparations, hybrid peptides, drug combinations, etc., can solve the problem that tumor cells evade the immune system and DCs cannot effectively present Tumor cells, etc.
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Embodiment 1
[0024] Example 1, Preparation of cell membrane lipids displaying GPI-NY-ESO-1 fusion protein on the surface
[0025] 1. Construction of recombinant plasmids and recombinant cells
[0026] 1. Insert the double-stranded DNA molecule (IgGFc-GPI fragment, 1374bp) shown in Sequence 3 of the sequence listing between the NheI and NotI restriction sites of the plasmid pCI-neo to obtain the recombinant plasmid pCI-pr-GPI.
[0027] 2. Insert the double-stranded DNA molecule shown in Sequence 2 of the sequence listing between the EcoRV and NheI restriction sites of the recombinant plasmid pCI-pr-GPI to obtain the recombinant plasmid pCI-pr-GPI / NY-ESO-1. According to the sequencing results, the structure of the recombinant plasmid pCI-pr-GPI / NY-ESO-1 is described as follows: a double strand shown in sequence 4 of the sequence table is inserted between the EcoRV and NotI restriction sites of the plasmid pCI-neo DNA molecule. The double-stranded DNA molecule shown in sequence 4 of the seq...
Embodiment 2
[0049] Example 2, Application of Cell Membrane Lipid Displaying GPI-NY-ESO-1 Fusion Protein on Surface and Cell Membrane Lipid Displaying GPI-NY-ESO-1 Fusion Protein on Surface with Oxidized Mannan Modification
[0050] Medium A-I: serum-free RPMI-1640 culture medium containing 30% (volume ratio) solution A-I.
[0051] Medium A-II: serum-free RPMI-1640 culture medium containing 30% (volume ratio) solution A-II.
[0052] Medium B-I: Serum-free RPMI-1640 culture medium containing 30% (volume ratio) Solution B-I.
[0053] Medium B-II: serum-free RPMI-1640 culture medium containing 30% (volume ratio) solution B-II.
[0054] Medium C (control medium): serum-free RPMI-1640 culture medium.
[0055] Medium D (control medium): Serum-free RPMI-1640 medium containing 50 μg / ml NY-ESO-1 polypeptide and 5 μl / ml Liposome 2000 (Invitrogen, product number 11668-019).
[0056] 1. Cell membrane lipids containing GPI-NY-ESO-1 fusion protein and DC cells promote human T lymphocytes to secrete c...
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