36 results about "Constant region gene" patented technology

Described herein are antibodies that bind to human tumor necrosis factor alpha (hTNFα). And list the nucleotide sequence of antibody heavy chain and light chain variable region and its derived amino acid sequence. The heavy chain and light chain variable region genes are respectively connected with the human immunoglobulin gamma 1 (hIgG1) heavy chain constant region and human kappa (k) light chain constant region genes to form a chimeric gene. Then, the vector containing the chimeric gene is introduced into the host cell line to express the antibody protein. This recombinant chimeric antibody protein can neutralize the activity of hTNFα in vitro, and is suitable for treating patients with excessive secretion of harmful hTNFα, including certain inflammations, such as rheumatoid arthritis and Crohn's disease.

The invention discloses an FPV (feline parvovirus)-resistant cat-derived genetic engineering antibody, and belongs to the technical field of antibody engineering. The antibody comprises a mouse-derived variable region and a cat-derived constant region, the nucleotide sequence of a heavy chain variable region of the antibody is represented as SEQ ID NO.1, the gene sequence of a heavy chain constant region of the antibody is represented as SEQ ID NO.5, the nucleotide sequence of a light chain variable region of the antibody is represented as SEQ ID NO.2, and the gene sequence of a light chain constant region of the antibody is represented as SEQ ID NO.6. The variable region sequence of a monoclonal antibody of a canine parvovirus is assembled with the cat-derived constant region, and the FPV-resistant cat-derived genetic engineering antibody is obtained, shows good FPV neutralization activity, has an effect of inhibiting red cell agglutination of the FPV, can be applied to the fields of cat-derived research for a canine parvovirus monoclonal antibody and the like and has great significance for promoting development of the cat-derived monoclonal antibody drugs.

The invention provides a method for preparing a pre-staining luminescent protein marker. The method comprises the following steps: acquiring a protein A gene, a protein G gene, an MBP (Myelin Basic Protein) gene and a heavy chain constant region gene (CH gene) of a mouse and rabbit derived antibody, and cutting and combining the genes according to sizes of different proteins in a pre-staining luminescent protein marker; expressing the proteins, purifying, and mixing proteins of different molecule weights according to a certain ratio so as to obtain a luminescent protein marker; mixing the prepared luminescent protein marker with the pre-staining luminescent protein marker according to different ratios. The pre-staining luminescent protein marker prepared by using the method is capable of recognizing primary antibodies or secondary antibodies of different sources, binding sites of proteins are increased, light signals are intensified, not only are general positions of proteins indicated in the electrophoresis process and after membrane transfer, but also precise positions of the proteins can be indicated on X-ray films, no extra pre-staining marker channels are needed, no extra antibodies are needed, and the luminescent proteins can be combined with IgG or anti-rabbit and anti-mouse secondary antibodies derived from species such as human beings, rats, mice and rabbits.

A target fused DNA vaccine pGJA-P for preventing decayed tooth is prepared by using the PCR process through amplifying the antigen 4 signal peptide of toxic T lymphocyte, the sequence of extracellular region, and the genes in Ig gamma 1 hinge region and constant region, respectively cloning them to pU cm-T carrier to configune pGTLA and pJIg, linking them to configure their recombinant plasmid, verifying the correctness of inserted fragments, and inserting the fusion gene in the front end of pGLUA-P. Its advantages are simple process, high safety, low cost, and high effect to improve immunity.

The invention discloses a humanized antibody expression vector and a construction method thereof. The construction method comprises the steps of: respectively connecting a 2A sequence-containing light chain constant region gene sequence CL-2A and a heavy chain constant region gene sequence CH with a pMD18-T vector to obtain a pMD18-T-CL-2A plasmid and a pMD18-T-CH plasmid; connecting the pMD18-T-CL-2A plasmid and a eukaryotic expression vector after being subjected to double digestion to obtain a CL-2A-containing eukaryotic expression vector; and then connecting the CL-2A-containing eukaryotic expression vector with a pMD18-T-CH plasmid to obtain a CL-2A-CH-containing eukaryotic expression vector. The construction method of the expression vector disclosed by the invention can be used for rapidly constructing a full-length complete antibody sequence only by inserting a variable region sequence of a mouse or a humanized antibody into a corresponding locus and rapidly constructing a chimeric antibody, the humanized antibody and the like, and is simple and practical.

The invention relates to an anti-HIV gene engineering recombinant virus and a preparation method thereof, and an anti-HIV gene engineering medicament. The anti-HIV gene engineering recombinant virus comprises a constant region of a human antibody, and a cluster of differentiation 4 (CD4) and a chemokine receptor 5 (CCR5) which are connected with the constant region. The preparation method for the anti-HIV gene engineering recombinant virus comprises the steps as follows: obtaining a heavy chain constant region gene of the human antibody, a light chain constant region gene of the human antibody, a CD4 gene and a CCR5 gene; constructing a virus vector shuttle plasmid comprising the four genes; co-transforming the shuttle plasmid and a virus auxiliary plasmid to generate a recombinant virus plasmid; and transferring the recombinant virus plasmid to a cell strain for culturing and purifying the virus. The recombinant virus has both the CD4 and CCR5 capable of specifically binding with the CD4 site and CCR5 site of an HIV virus, an obviously enforced specific binding effect, and the function of doubly preventing the HIV virus from infecting a host cell. The preparation method is simple and practical. The anti-HIV gene engineering medicament with the recombinant virus can effectively act on the HIV virus, and can further effectively prevent and treat the infection of the HIV virus.

The invention provides a bifunctional antibody and a construction method thereof. The bifunctional antibody comprises an antibody heavy chain constant area, an antibody light chain constant area, a first target protein, which is connected to the antibody heavy chain constant area, and a second target protein, which is connected to the antibody light chain constant area. The construction method of the bifunctional antibody comprises the following steps: individually obtaining the antibody heavy chain constant area gene, antibody light chain constant area gene, the first target gene, and the second target gene; adopting expression carriers to construct recombinant carrier shuttle plasmids of the genes mentioned above; transfecting the recombinant carrier shuttle plasmids to an expression bacterium strain to carry out culture so as to obtain the bifunctional antibody; carrying out centrifugation, and collecting the supernate or purifying the cells so as to obtain the bifunctional antibody. The construction method assembles antibody molecules in a gene level, thus the pertinence is strong, and the method is easy to achieve. The obtain bifunctional antibody comprises two antigen combining sites, so the bifunctional antibody can combine two different antigens at the same time.

The invention discloses a variable region sequence of a specific anti-chlorothalonil antibody. The amino acid sequence of a heavy chain variable region coding gene is shown in SEQ ID NO: 2. The invention also discloses an anti-chlorothalonil recombinant full-length IgG antibody. The invention also discloses a recombinant antibody expression plasmid. The heavy chain variable region and light chain variable region sequences of the anti-chlorothalonil recombinant full-length IgG antibody are derived from a monoclonal cell strain which secretes a high-affinity and high-specificity anti-chlorothalonil antibody and is obtained by immunizing mice for multiple times, performing cell fusion and screening. The sequence genes are respectively connected to expression vectors containing a mouse IgG1 heavy chain constant region gene and a mouse kappa light chain constant region gene, the recombinant full-length IgG antibody is obtained by expression of mammalian cells of a double-plasmid system, the recognition activity of the recombinant full-length IgG antibody is very close to that of a mouse-derived parent antibody, large-scale standardized production of the anti-chlorothalonil antibody can be realized, and reliable core raw materials are provided for various immunoassay methods for rapid detection of chlorothalonil residues.

The invention discloses a variable region sequence of a specific anti-thiacloprid antibody. The amino acid sequence of a heavy chain variable region encoding gene is disclosed by SEQ ID NO: 2. The invention also discloses an anti-thiacloprid recombinant overall-length antibody, which comprises a heavy chain constant region, a heavy chain variable region, a light chain constant region and a light chain variable region, wherein the amino acid sequence of the heavy chain variable region encoding gene is disclosed by SEQ ID NO: 2. The invention also discloses an antibody expression plasmid. A sequence gene obtained by the invention is independently connected to expression carriers containing a heavy chain constant region gene and a light chain constant region gene, mammal cell of a double-plasmid system is adopted to obtain the recombinant overall-length antibody, the identification activity of a murine parent antibody is guaranteed, the anti-thiacloprid antibody can be stably produced ona large scale, and a reliable core reagent is provided for various immunoassay methods for detecting thiacloprid residues.

The invention relates to a single chain antibody of human anti-placenta growth factor, which is encoded by a heavy-chain gene with an SEQ NO.1 sequence and light-chain gene with an SEQ NO.2 sequence.The invention also provides a preparation method thereof, comprising the following steps in sequence: amplifying human total RNA heavy-chain variable region, light-chain variable region and constant region gene of the whole set by a PCR technique, splicing and constructing an overall-length single-chain antibody cDNA library; expressing the cDNA library by an externally-coupled transcription/translation system to obtain an antibody-ribosome-mRNA compound; performing affiliation selection and clution on the compound by a magnetic bead coated by specific antigen; amplifying selected destinationantibody gene by the PCR technique; and expressing the amplified antibody gene using the externally-coupled transcription/translation system repeatedly to obtain the single-chain antibody. The antibody is adapted to prepare medicament for treating ovarian cancer.

The invention discloses a preparation method of an allergen-specific IgE antibody composite quality control product and the allergen-specific IgE antibody composite quality control product. The preparation method specifically comprises the following steps: preparing an allergen-specific IgE antibody; obtaining a target gene, and obtaining a constant region gene sequence and a variable region gene sequence of human IgE; the method comprises the following steps: designing a primer containing an XhoI restriction enzyme cutting site according to a heavy chain constant region gene sequence of human IgE, designing a primer containing an Hind III restriction enzyme cutting site according to a light chain constant region gene sequence of human IgE, and constructing an expression empty vector containing a light chain signal peptide, a heavy chain signal peptide, a human IgE heavy chain and a light chain C region gene fragment; constructing recombinant expression vectors of different allergens; transfecting the cells; cloning, recognizing and screening; the quality control product has the advantages of high titer, higher purity, favorable dilution linearity and favorable stability, and has higher use value in the aspects of quality control, conventional quality control, performance verification and product development of large-scale production.

The invention discloses a variable region sequence of a specific anti-thiacloprid antibody. The amino acid sequence of the gene encoding the heavy chain variable region is shown in SEQ ID NO:2. The present invention also discloses an anti-thiacloprid recombinant complete antibody, including heavy chain constant region, heavy chain variable region, light chain constant region and light chain variable region, the amino acid sequence of the gene encoding the heavy chain variable region is as follows: Shown in SEQ ID NO:2. The invention also discloses an antibody expression plasmid. The sequence genes obtained in the present invention are respectively connected to the expression vectors containing the heavy chain constant region gene and the light chain constant region gene, and the recombinant complete antibody is obtained by using the mammalian cell expression of the double plasmid system, which ensures the recognition of the mouse parent antibody activity, and enables the stable production of anti-thiacloprid antibodies in large quantities, and provides reliable core reagents for various immunoassay methods for the detection of thiacloprid residues.

The invention relates to a recombinant chimeric Fab antibody comprising a recombinant Fd and a recombinant chimeric light chain which binds to hepatitis B surface antigen with high affinity, generated by fusing variable region genes (V_H and V_L) of an anti-HBsAg mouse antibody 5S and constant region genes of human, CH1 region of human IgGI and the CL region of human kappa chain, wherein mouse V_L and human CL are linked by overlap PCR to generate chimeric light chain and mouse V_H and CH₁ are linked by overlap PCR to generate chimeric Fd.