Human immunodeficiency virus affinity adsorption column, and preparation method and uses thereof
An HIV and adsorption column technology is applied in the field of rapid and efficient removal of HIV from blood, which can solve the problems of ineffective immunotherapy, difficult opportunistic treatment, frequent antiviral treatment effects, etc. The effect of short treatment time and large toxic and side effects
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Embodiment 1
[0054] Cloning, expression and purification of embodiment 1 affinity protein
[0055] Affinity proteins include CD4 molecules, CCR-5, CXCR-4, and gp120 antibodies, all of which can be expressed by eukaryotic yeast expression systems. The following is an example of the main receptor CD4 molecule.
[0056] Human CD4 molecule is a kind of single-chain transmembrane glycoprotein with a relative molecular weight of about 55kDa, which belongs to the immunoglobulin superfamily and is mainly expressed in some T cells, thymocytes, some B cells and Epstein-Barr virus-transformed B cells, single Nuclei - Macrophages and the surface of brain cells in specific areas. It plays an important role in the development of T cells, the activation of mature T cells, and signal transduction. It is an important effector molecule in the occurrence of adaptive immune responses and plays a huge role in maintaining the normal physiological functions of the body.
[0057] The human CD4 molecular cDNA ...
Embodiment 2
[0148] The preparation of embodiment 2 immunoadsorption microspheres
[0149] principle:
[0150] The affinity microspheres can be glass microspheres with a size of 1 mm, chitosan cross-linked microspheres with a diameter greater than 500 μm, or agarose microspheres.
[0151] In this experiment, the choice of particle size can also choose 1mm glass beads as the carrier material. Due to their strong surface area and pore structure, glass beads have the potential to immobilize various ligands. Moreover, glass beads also have relatively good blood compatibility, so they can also be considered as a carrier for adsorption. Before protein immobilization, glass beads need to be pretreated such as washing. During the preparation process, glass beads are often mixed with impurities such as dust and grease, so they need to be cleaned. In this experiment, the glass beads were placed in a solution of 95% acetone and ultrasonicated for 5 minutes to remove impurities from the glass be...
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