204 results about "Hybridoma technology" patented technology

The invention relates to a competitive ELISA method based on a foot-and-mouth disease A type VP1 protein and its monoclonal antibody, also relates to a preparation method of the foot-and-mouth disease A type VP1 protein, and a preparation method of the monoclonal antibody of the foot-and-mouth disease A type VP1 protein, and belongs to the technical field of animal immunological detection. In the invention, a primer pair C1 and C2 and a primer pair E1 and E2 are amplified to obtain a gene sequence of the foot-and-mouth disease A type VP1 protein, the foot-and-mouth disease A type VP1 protein is obtained by constructing an expression plasmid, introducing the expression plasmid into a prokaryotic expression host and carrying out inducible purification, the foot-and-mouth disease A type VP1 protein monoclonal antibody is obtained by treating the foot-and-mouth disease A type VP1 protein as an antigen through a hybridomas technology, and the competitive ELISA method used for detecting a foot-and-mouth disease A type antibody is established based on the foot-and-mouth disease A type VP1 protein and its monoclonal antibody. The detection method has a strong specificity and a good stability, and can be used for detecting a foot-and-mouth disease A type serum antibody. By comparing a result obtained through the detection method with a liquid phase blocking ELISA kit, the coincidence rate is 95.8%.

By use of hybridoma technology, recombinant human Delta like 4 (rhDll4) is used as an antigen for immunizing a BALB / c mice to obtain a high affinity and biological activity anti-human Delta like 4 monoclonal antibody. The monoclonal antibody is characterized in that: the monoclonal antibody can be combined with rhDll4 specifically, and can block human umbilical vein endothelial cells (HUVEC) proliferation suppression of the rhDll4. Specifically, screening, a preparation method, and nucleotide and amino acid sequences of the heavy chain variable region and the light chain variable region of the anti-human Delta like 4 monoclonal antibody are disclosed, and the nucleotide and amino acid sequences comprise nucleotide and amino acid sequences corresponding to complementarity determining regions CDR1, CDR2 and CDR3.

The invention relates to a humanized monoclonal antibody targeting Claudin18.2 as well as a preparation method and application thereof. According to the present invention, the Claudin18.2 humanized monoclonal antibody obtained through the cell fusion and the hybridoma technology can be highly specifically combined with CHO-Claudin18.2 cells, but is almost not combined with CHO-Claudin18.1 cells, the preparation method of the Claudin18.2 humanized monoclonal antibody of the present invention has simple steps, and the obtained Claudin18.2 humanized monoclonal antibody has good ADCC and CDC activity on the expression of Claudin18.2 positive tumor cells. The humanized monoclonal antibody of Claudin18.2 disclosed by the invention is high in specificity and small in side effect, and has a very good prospect in application to treatment and / or prevention or diagnosis of Claudin18.2-related diseases such as tumors.

The invention relates to a clostridium perfringens alpha toxin double-antibody sandwich ELIS quantitative determination method. According to the clostridium perfringens alpha toxin double-antibody sandwich ELIS quantitative determination method, alpha toxin protein obtained via prokaryotic expression is taken as an immunogen; monoclonal antibodies obtained via hybridoma technique are taken as detection antibodies and capture antibodies; reaction conditions are optimized via experiments; alpha toxin protein samples with a series of concentration are used for construction of a standard curve; the double-antibody sandwich ELIS method is established; and indexes of the double-antibody sandwich ELIS are verified. The clostridium perfringens alpha toxin double-antibody sandwich ELIS quantitative determination method specifically comprises following steps: (1) prokaryotic expression of alpha toxin; (2) preparation of anti-alpha toxin monoclonal antibodies; (3) establishment of the double-antibody sandwich ELIS method; (4) establishment of the standard curve; and (5) performance evaluation on the double-antibody sandwich ELIS method. The clostridium perfringens alpha toxin double-antibody sandwich ELIS quantitative determination method is excellent in specificity, and high in sensitivity and stability, is fast and convenient, and can be used for effective quantitative determination of alpha toxin in A-E type clostridium perfringens cultural supernatants; and the high-efficient detection method is provided for alpha toxin determination.

The invention relates to a mouse anti-human CIAPIN1 monoclonal antibody for confirming multi-drug resistant gastric cancer, and a preparation method thereof. The invention is characterized in that the monoclonal antibody is obtained by adopting the hybrid tumor technology; the heavy chain is 1gG1; the light chain is k; the molecular weight is 39KD; and the affinity is 3.6x10. The preparation method comprises the following steps of: (1) synthesizing primers; (2) synthesizing single-chain DNA by carrying out reverse transcription on SGC-7901 / VCR general RNA of a human gastric-cancer multi-drug resistant cell; (3) expanding human CIAPIN1 gene cDNA by using the primers F and R and by adopting a PCR method; (4) constructing CIAPIN1 gene induction expression plasmids pET28-CIAPIN1 and pGEX-CIAPIN1; (5) constructing engineering bacteria B21-CIAPIN1 and DE3-CIAPIN1; (6) using the engineering bacteria B21-CIAPIN1 and DE3-CIAPIN1 to carry out prokaryotic expression and purification on CIAPIN1 protein; (7) using CIAPIN1 protein to immunize a 4-week-old Balb / C mouse so that mouse anti-human CIAPIN1 antibody is generated; and (8) using the serum of the immunized mouse to prepare the mouse anti-human CIAPIN1 monoclonal antibody. The invention can better identify multi-drug resistant gastric cancer and non-multi-drug resistant gastric cancer of clinical gastric cancer patients, and predict clinical prognosis of the gastric cancer patients.

The invention discloses human-mouse chimeric Siglec-15-resistant neutralizing full-molecule IgG and a preparation method and application thereof, and belongs to the field of biological pharmacy. Miceimmune to Siglec-15-specific peptide are adopted, a mouse-derived Siglec-15-resistant monoclonal antibody is prepared through a hybridoma technology, and the recombinant human-mouse chimeric Siglec-15-resistant full-molecule IgG is prepared by using a genetic engineering technology and an antibody engineering technology. Specific amino acid segments in the extracellular region of Siglec-15 can berecognized effectively through the chimeric antibody, and combination of Siglec-15 protein with Sialyl-Tn protein is inhibited. The invention discloses the human-mouse chimeric Siglec-15-resistant full-molecule IgG, the human-mouse chimeric Siglec-15-resistant full-molecule IgG includes a light chain variable region and a heavy chain variable region, wherein the amino acid sequence of the light chain variable region is shown in SEQ ID NO. 5, and the amino acid sequence of the heavy chain variable region is shown in SEQ ID NO. 6 or a conservative variant which is obtained through conservative mutation of the amino acid sequence by using addition, deletion, substitution and modification of one or more amino acids. The invention also discloses a DNA molecule, expression vector, host cells andapplication of the full-molecule IgG antibody.

The invention relates to a method for detecting the residual immunity of an aryl-N-methylcarbamates pesticide, and belongs to the biotechnology field. The method includes the steps as follows: a carrier protein is coupled with a general hapten (HA) of the aryl-N-methylcarbamates pesticide to obtain a complete antigen; by applying the hybridoma technique, the antigen immunized Balb / C mice are screened by adopting the ELISA method and subcloned by adopting the limiting dilution method to obtain the hybridoma cell strains capable of stably secreting the aryl-N-methylcarbamates pesticide; and monoclonal antibodies are prepared. Based on the monoclonal antibodies, four ELISA detection methods for detecting the residual immunity of the aryl-N-methylcarbamates pesticide are established. The method has high specificity, sensitivity and stability, and provides a quick, sensitive and specific technology for trace-detecting the aryl-N-methylcarbamates pesticides.