59 results about "Epithelial cell adhesion molecule" patented technology

The invention discloses a CTC (circulating tumor cell) capturing and purifying substrate based on chitosan nanoparticles as well as a preparation method and an application of the CTC capturing and purifying substrate. Chitosan with good cytocompatibility is selected for constructing a three-dimensional nanostructure 'soft' substrate, anti-adhesion molecules such as PEG (polyethylene glycol) molecules are introduced to reduce the nonspecific adhesion of cells on an interface, affine CTC capturing molecules such as EpCAM (epithelial cell adhesion molecule) aptamers are used for high-specific capturing of CTCs, and the purity of target cells is further improved in an in-situ culturing manner of the captured cells on the basis of proliferation attribute difference of the CTCs and blood cells. The CTC capturing and purifying substrate has good cytocompatibility and can keep the activity of the captured cells, a preparation process is simple and convenient, the cost is low, in-situ culturing and purification of the captured cells can be realized, and large-scale implementation is easy to realize.

The invention provides an aptamer EpCAM (epithelial cell adhesion molecule) Ccut of an EpCAM as well as a preparation method and application thereof, relating to nucleic acids. The aptamer has high specificity and affinity. The aptamer has a G tetramer structure or a stem-loop structure. The preparation method comprises the steps of designing and synthesizing a single-stranded DNA (deoxyribonucleic acid) random oligonucleotide bank, screening a target oligonucleotide sequence, and identifying the target protein binding specificity and affinity of the target oligonucleotide sequence by a flow analysis method. The screened aptamer does not have toxicity, has small molecular weight and good permeability, is easy to synthesize and label, only specifically identifies EpCAM proteins and cell lines expressing the EpCAM proteins, and does not have a function of identifying the cell lines which do not express the proteins.

The invention discloses joint application of three types of monoclonal antibody-coupled immunomagnetic beads to sorting tumor cells and a method for sorting breast cancer tumor cells. The three types of monoclonal antibody-coupled immunomagnetic beads include anti-HLA-G (human leukocyte antigen-G) monoclonal antibody-coupled immunomagnetic beads, anti-EpCAM (epithelial cell adhesion molecule) monoclonal antibody-coupled immunomagnetic beads and anti-CK8 / 18 (casein kinase 8 / 18) monoclonal antibody-coupled immunomagnetic beads. The joint application and the method have the advantages that the three types of jointly applied antibody-coupled immunomagnetic beads are high in breast cancer tumor cell enrichment ratio and good in sensitivity and repeatability, and the relevant tumor cells can be detected; the three types of immunomagnetic beads (the anti-EpCAM immunomagnetic beads, the anti-HLA-G immunomagnetic beads and the anti-CK8 / 18 immunomagnetic beads) are jointly applied to enriching the tumor cells, so that the circulating tumor cell coverage rate can be increased to the greatest extent, the enrichment ratio can reach 80%, and a technology is international pioneer; the tumor cell enrichment specificity and sensitivity can be particularly guaranteed owing to usage of the anti-HLA-G monoclonal antibody-coupled immunomagnetic beads.

The invention relates to a nucleic acid, in particular to a nucleic acid aptamer of the epithelial cell adhesion molecule and a preparation method thereof. The invention provides a high-specificity and high-affinity nucleic acid aptamer of the epithelial cell adhesion molecule EpCAM and a preparation method and applications thereof. The nucleic acid aptamer of the epithelial cell adhesion molecule EpCAM is of the G-quadruplex structure or the stem-loop structure. The preparation method comprises the following steps: designing and synthetizing a single-stranded DNA (deoxyribonucleic acid) random oligonucleotide library, screening the target oligonucleotide sequence, and identifying the binding specificity and affinity of the sequence to the target protein through the flow cytometry. The screened nucleic acid aptamer is non-toxic, has small molecular weight and good permeability, is easy in synthesis and marking and can only perform specific identification to the EpCAM protein and perform specific identification and specific expression to the cell line of the EpCAM protein; and the nucleic acid aptamer can not identify the cell line which does not express the protein.

The invention discloses a preparation method of EpCAM (Epithelial Cell Adhesion Molecule) antibody immunomagnetic beads. The preparation method comprises the following specific steps: synthesizing oleic acid-Fe3O4; preparing Fe3O4 / PVAc (Polyvinyl Acetate) magnetic microspheres; preparing (Fe3O4 / PVA) / SiO2 magnetic microspheres; adding aminated magnetic nanoparticles into a PBS (Polybutylene Succinate) solution of glutaraldehyde and stirring for 3h; modifying an aldehyde-group functional group on the surfaces of the magnetic particles through a Schiff base formed between an amino group and an aldehyde group; after finishing reaction, separating out the particles from a reaction medium by utilizing an external magnetic field and dispersing the particles into the PBS solution; carrying out hydrazino modification on an antibody: adding a dimethylformamide solution containing p-hydrazone pyridine formic acid N-hydroxysuccinyl imide into an EpCAM antibody solution; after reacting for 2h, purifying by utilizing an ultra-filtration column to obtain a hydrazino-modified EpCAM antibody; preparing the immunomagnetic beads: mixing the aldehyde-group-modified magnetic microspheres and the hydrazino modified EpCAM antibody and mixing to obtain the EpCAM antibody immunomagnetic beads of EpCAM antibody coupled magnetic microspheres. The EpCAM antibody immunomagnetic beads disclosed by the invention have the advantages of good stability, strong specificity and the like.

The invention provides an aptamer EpCAM (epithelial cell adhesion molecule) B of an EpCAM as well as a preparation method and application thereof, relating to nucleic acids. The aptamer has high specificity and affinity. The aptamer has a G tetramer structure or a stem-loop structure. The preparation method comprises the steps of designing and synthesizing a single-stranded DNA (deoxyribonucleic acid) random oligonucleotide bank, screening a target oligonucleotide sequence, and identifying the target protein binding specificity and affinity of the target oligonucleotide sequence by a flow analysis method. The screened aptamer does not have toxicity, has small molecular weight and good permeability, is easy to synthesize and label, only specifically identifies EpCAM proteins and cell lines expressing the EpCAM proteins, and does not have a function of identifying the cell lines which do not express the proteins.

The invention belongs to the field of molecular biology and particularly relates to EpCAM (epithelial cell adhesion molecule) single-domain antibody E6. A single-domain heavy-chain antibody library capable of specifically binding with target antigen EpCAM is screened from a camelid source immunized single-domain heavy-chain antibody library by means of a phage display technique; a gene sequence toencode the antibody is cloned to a prokaryotic expression vector so as to obtain the efficient-expression EpCAM single-domain antibody. The specific binding performance of the single-domain antibodycan be significantly improved, and basis is laid for immuno-guided clinical therapy of EpCAM.

The invention provides an aptamer EpCAM (epithelial cell adhesion molecule) D of an EpCAM as well as a preparation method and application thereof, relating to nucleic acids. The aptamer has high specificity and affinity. The aptamer has a G tetramer structure or a stem-loop structure. The preparation method comprises the steps of designing and synthesizing a single-stranded DNA (deoxyribonucleic acid) random oligonucleotide bank, screening a target oligonucleotide sequence, and identifying the target protein binding specificity and affinity of the target oligonucleotide sequence by a flow analysis method. The screened aptamer does not have toxicity, has small molecular weight and good permeability, is easy to synthesize and label, only specifically identifies EpCAM proteins and cell lines expressing the EpCAM proteins, and does not have a function of identifying the cell lines which do not express the proteins.