Compositions and methods for preparing t cell compositions and uses thereof
A cell and cancer cell technology, applied in biochemical equipment and methods, drug combinations, animal cells, etc., can solve inherently complex and cumbersome problems
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Embodiment 1
[0513] Example 1. Accurate Neostim Clinical Process
[0514] In this paper, late T cell therapy is provided, in which the pre-verified preserved subject cancer-specific antigen peptide pre-sensitive and responded T cells is applied to the subject. The method provided in this embodiment avoids lengthy sequencing, identifying and producing a subject-specific novel antigen peptide during the time of such evaluation and preparation of the subject, and the preparation of the preparation of the subject. The T cells having a subject specific TCR for cancer immunotherapy were produced. The advantage of this method is its rapid, targeted, and stable. like Figure 1A As shown in, patients who have been identified having cancer or tumors can be administered T cells, and the T cell is activated by peptide-managed peptide, which has a peptide-managed peptide having cancer known to identify. The selection of epitopes produced by the library of shared antigens, pre-verified collection. Process o...
Embodiment 2
[0515] Example 2. Production of target tumor cell antigen response T cells in vitro
[0516] Material:
[0517] AIM V medium (INVITROGEN)
[0518] Human flt3l, pre-clinical Cellgemx # 1415-050 Reserve 50NG / μL
[0519] TNF-α, preclinical Cellgenix # 1406-050, reserve 10 ng / μL
[0520] IL-1β, preclinical Cellgenix # 1411-050, reserve 10 ng / ul
[0521] PGE1 or Alprostadil-Cayman (from Czech Republic) Reserve 0.5μg / μL
[0522] R10 medium-rpmi 1640glutamax + 10% human serum + 1% PenStrep
[0523] 20 / 80 medium -18% AIM V + 72% rpmi 1640GLUTAMAX + 10% serum + 1% PenStrep
[0524] IL7 reserve 5NG L
[0525] IL15 reserve 5 ng / μL
[0526] program:
[0527] Step 1: In a 2 ml AIM V medium, 5 million PBMC (or target cells) is paved in each hole of 24-well plates with FLT3L.
[0528] Step 2: Peptide load and mature in AIMV
[0529] 1. Mix the target peptide library (except for peptide conditions) with PBMC (or target cells) in each well.
[0530] 2. Cost for 1 hour.
[0531] 3. Afte...
Embodiment 3
[0550] Example 3. Evaluation of antigen's presence
[0551] For a subset of the predicted antigen, the affinity of the new epitope and a new epitope having an HLA allele is determined for the HLA allele. Stability. Exemplary data of the subset of the RAS new antigen and GATA3 new antigen is shown Figure 4A and Figure 20 middle.
[0552] Exemplary Detailed Description of Scheme for Measuring Binding Prolycity of Peptide and I MHC has been announced (SETTE et al., Mol.immunol. 31 (11): 813-22, 1994). Briefly, the MHCI complex is prepared and combined to the radiolabeled reference peptide. The peptide was incubated with these complexs with different concentrations with these complexes and the remaining amount of the peptide bonded to MHCI was measured using a dimensional exclusion gel filtration. The lower the concentration of the test peptide required for the replacement of the radioactive labeled reference peptide indicates the more affinity of the test peptide to MHCI. Peptides o...
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