8 results about "Freunds complete adjuvant" patented technology

The invention belongs to the field of antibody development, and discloses a screening method and application of a human-mouse protein high homologous target antibody based on a fully humanized antibody mouse. Aiming at the problem of weak antibody response caused by immune tolerance of human-mouse high homologous targets (protein homology is greater than or equal to 95%), the following scheme is provided: in embryonic stem cells (ES cells) of HUGO-Mabfully humanized antibody transgenic mice, a mouse target gene (such as ACVR2A) is knocked out through a Turbo Knockout technology, and homozygous knockout ES clones are screened; carrying out microinjection on the clones to the whitened B6 mouse blastocyst, and transplanting a pregnant mouse to obtain a Founder mouse; the Founder mouse is subjected to target antigen immunization for more than or equal to 4 times (the Freund's complete adjuvant is used for the first time), and the titer of the serum antibody is detected. According to the invention, 100% homozygous knockout chimeric efficiency is realized in the Founder stage, the mouse construction period is shortened from traditional 8-10 months to 3-4 months, and the diversity and affinity of the antibody are significantly improved (titer reaches 1: 729,000). The obtained antibody can be used for preparing medicines for treating tumors or autoimmune diseases.

The application discloses a SjHNF4 polypeptide antibody, a preparation method and application. The preparation method comprises the following steps: downloading a SjHNF4 protein sequence, screening a polypeptide sequence, adding a cysteine at an N terminal, then performing polypeptide synthesis and purification, coupling to obtain a recombinant polypeptide, mixing the coupled polypeptide with Freund's complete adjuvant to obtain a mixture, immunizing a mammal by using the mixture, taking heart blood of the mammal and separating serum to obtain a polypeptide antibody. The polypeptide antibody is detected by using an ELISA method, and then the polypeptide antibody is purified by using a polypeptide affinity chromatographic column. Finally, the binding affinity of the antibody and the antigen is detected by Western Blot and immunofluorescence experiments. The polypeptide antibody obtained by the application has high purity (not less than 90%) and high titer (not less than 512000), and has good purity and immunogenicity, can satisfy various experimental requirements and has commercial value.

The present application relates to the biomedical technology field, disclose a kind of dry syndrome intestinal damage mouse model construction method and its application.The construction method includes: (1) preparation contains Freund's complete adjuvant and 1mg / mL mouse intestinal epithelial cell antigen immunization emulsifier I and preparation contains Freund's incomplete adjuvant and 0.5mg / mL mouse intestinal epithelial cell antigen immunization emulsifier II;(2) to mouse injection the immunization emulsifier I or the immunization emulsifier II in batches, obtain the dry syndrome intestinal damage mouse model.The dry syndrome intestinal damage mouse model construction method provided by the present application only immunizes mouse by mouse intestinal epithelial cell antigen and adjuvant, obtains the mouse model that simultaneously shows dry syndrome and intestinal damage.The construction method not only has the advantages of high efficiency, fast, economic, but also is extremely easy to popularize and widely used, has good application value.

The invention belongs to the technical field of biological detection, and particularly relates to a monoclonal antibody for HPV16 type E7 protein detection and application of the monoclonal antibody. E7 protein serving as an antigen and a Freund's complete adjuvant are mixed, then a mouse is subjected to primary immunization, then the E7 protein and a Freund's incomplete adjuvant are mixed, then the mouse is subjected to enhanced immunization, and the HPV16 type E7 protein detection monoclonal antibody is obtained. The monoclonal antibody for detecting the HPV16 type E7 protein is prepared by taking mouse splenocytes and myeloma cells to be fused to prepare hybridoma cells, then performing clone detection on the hybridoma cells to screen the hybridoma cells aiming at the specificity of the HPV16 type E7 protein, and finally obtaining the monoclonal antibody for detecting the HPV16 type E7 protein. According to the present invention, the HPV16 type E7 antigen detection kit is developed by using the immunochromatography method, and the kit has advantages of high sensitivity, strong specificity, simple operation and fast detection speed, and can be used for rapid screening diagnosis and illness state monitoring of HPV16.

The application belongs to the technical field of medicine. The monkeypox virus surface membrane antigen H3L protein expressed by Escherichia coli is used as an antigen, and Balb / c mice are inoculated with the H3L protein emulsified with Freund's complete adjuvant, and the antigen is co-immunized for three times. The mouse spleen cells are fused with myeloma cells SP2 / 0 cells to prepare H3L specific hybridoma cells, and then the hybridoma is cloned and detected to screen the hybridoma cells specific to the H3L antibody. After three rounds of antigen specific screening, a high-affinity poxvirus murine monoclonal antibody is obtained, the amino acid sequence of the heavy chain variable region domain is shown as SEQ ID NO. 1 in the sequence table, and the amino acid sequence of the light chain variable region domain is shown as SEQ ID NO. 3 in the sequence table. The antibody has good binding capacity with the antigen H3L through ELISA detection, and has good protection in the in vitro neutralization experiment. The antibody can not only be applied to the research and treatment of diseases caused by poxvirus infection, but also can be used as a tool for poxvirus detection and treatment, and has potential application value in detecting poxvirus infection.

The invention relates to the technical field of biomedicine, and discloses a construction method and application of a sicca syndrome intestinal damage mouse model. The construction method comprises the following steps: (1) preparing an immune emulsifier I containing a Freund's complete adjuvant and 1mg / mL of mouse intestinal epithelial cell antigen, and preparing an immune emulsifier II containing a Freund's incomplete adjuvant and 0.5 mg / mL of mouse intestinal epithelial cell antigen; and (2) injecting the immune emulsifier I or the immune emulsifier II into a mouse for several times to obtain the sicca syndrome intestinal damage mouse model. According to the construction method of the sicca syndrome intestinal damage mouse model, the mouse is immunized only through the mouse intestinal epithelial cell antigen and the adjuvant, and the mouse model with the sicca syndrome and the intestinal damage at the same time is obtained. The construction method not only has the advantages of high efficiency, rapidness and economy, but also is very easy to popularize and widely use, and has very good application value.

The application discloses a specific antibody targeting a lysine K1106 site of an ITSN1-S protein lactylation and a preparation method and application thereof, and the preparation method comprises the following steps: mixing and emulsifying an ITSN1-S-K1106 lactylation antigen with Freund's complete adjuvant and Freund's incomplete adjuvant respectively to obtain a first immunization liquid drop and a booster immunization liquid drop, performing animal immunization by using the first immunization liquid drop and the booster immunization liquid drop respectively, and separating the specific antibody targeting the lysine K1106 site of the ITSN1-S protein lactylation from blood of the immunized animal.The antibody provided by the application can specifically detect the lactylation modification condition of the human ITSN1-S protein from the cell and tissue levels in actual application, and is helpful to explore the action mechanism of post-translational modification of the ITSN1-S protein in tumor proliferation, migration and the like, and provides a potential action target for clinical diagnosis and treatment of tumor diseases.

This invention utilizes an insect-baculovirus expression system to prepare the recombinant protein SADS-CoV-S of porcine acute diarrhea syndrome coronavirus, which is then used to prepare egg yolk antibodies. By optimizing the composition of the subunit vaccine composition for immunizing laying hens, IL-2 and ZnCl2 solution are added as immunostimulants to the conventional adjuvant, increasing the antibody concentration in the egg yolk antibody extract and improving the neutralizing antibody titer and protective efficacy. When IL-2 and zinc chloride aqueous solution are added to the immunization composition simultaneously, the resulting egg yolk antibody aqueous solution exhibits the highest protein concentration, and at the same protein concentration, the egg yolk antibody has the highest neutralizing titer. Compared to the immunization composition using Freund's complete adjuvant alone, the neutralizing titer is increased by more than 60 times.