Preparation method and application of pMHC polymer based on DNA nanotechnology
A nanotechnology and institutional technology, applied in the field of T cell detection, can solve the problems of insufficient binding affinity and unsatisfactory performance, and achieve the effect of cost saving and high efficiency
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Embodiment 1
[0054] Example 1 PMHC multi -gathering system preparation method based on DNA nanotechnology
[0055] This example provides a precise space assembly method and maximum assembly efficiency test based on DNA nano -technology control PMHC monomer.
[0056] First, SPYTAG, which is modified by clicked by clicking a chemical reaction to connect with the complementary chain containing DBCO, and use 100kda centric filters to purify the product.
[0057] Secondly, the SPYTAG-DNA complementary chain and PE modified complementary chain and rectangular DNA nano structure (with the corresponding capture chain) are annealing in the PCR instrument.And purified through 100kda centric filters to remove excess proteins.Among them, the preparation method with a rectangular DNA nanogenic structure with a specific capture chain is as follows: 1 template chain and several short chains are paired with a complementary pairing of alkali base to form a rectangular DNA nano structure with a specific capture...
Embodiment 2
[0070] Example 2 peptide based on DNA nanotechnology-Main tissue compatibility complex (DNA-PMHC) polyette detection and analysis of specific T cell efficiency assessment
[0071] In order to test the detection efficiency of the specific T cells of the present invention, C57BL / 6 mice spleen cells were selected for testing.
[0072] First, use OVA 257-264 (SIINFEKL) antigen peptides and Polyicica (IC) mixture to 1: 1 subcutaneous immune C57BL / 6 mice, immune every 3 days for immune 3 times.After 7 days, remove the spleen and prepare the spleen single -cell suspension.
[0073] Second, use H-2K b -OVA 257-264 (SIINFEKL) Teti-cluster as a control and DNA nanogenic polymerizer for dyeing H-2K, respectively b Limited OVA257-264 specific TCR.
[0074] Finally, use the current cytometer 2060R, Aglen) to measure the sample, and use Novoexpress to analyze data.
[0075] The experimental results show that the use of DNA-PMHC to detect antigen-specific T cells. The number of PMHCs from 3-12 D...
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