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94 results about "Synthetic DNA" patented technology

Process for producing humanized chimera antibody

A process for the production of humanized chimera antibody, wherein the chimera antibody is produced easily without changing any of the amino acids of its mouse antibody variable region, which comprises the steps of: (1) constructing a cassette vector by inserting a cDNA coding for a heavy chain constant region of human antibody into an expression vector for animal cell use and establishing a cloning site in the upstream region of the heavy chain constant region of said cassette vector for inserting a cDNA which encodes a heavy chain variable region of nonhuman animal antibody; (2) digesting a cDNA coding for the heavy chain variable region of nonhuman animal antibody with restriction enzymes; (3) inserting said cDNA coding for the heavy chain variable region of nonhuman animal antibody into the cassette vector, using a synthetic DNA which comprises a base sequence corresponding to the 5'-end side of said heavy chain constant region of human antibody and a base sequence corresponding to the 3'-end side of said heavy chain variable region of nonhuman animal antibody and is possessed of the restriction enzyme recognition sites on both of its ends, thereby constructing a humanized chimera antibody heavy chain expression vector in which said cDNA coding for the heavy chain constant region of human antibody and said cDNA coding for the heavy chain variable region of nonhuman animal antibody are linked together through said synthetic DNA; (4) constructing a cassette vector by inserting a cDNA coding for a light chain constant region of human antibody into an expression vector for animal cell use and establishing a cloning site in the upstream region of the light chain constant region of said cassette vector for inserting a cDNA which encodes a light chain variable region of nonhuman animal antibody; (5) digesting a cDNA coding for the light chain variable region of nonhuman animal antibody with restriction enzymes; (6) inserting said cDNA coding for a light chain variable region of nonhuman animal antibody into the cassette vector using a synthetic DNA which comprises a base sequence corresponding to the 5'-end side of said light chain constant region of human antibody and a base sequence corresponding to the 3'-end side of said light chain variable region of nonhuman animal antibody and is possessed of the restriction enzyme recognition sites on both of its ends, thereby constructing a humanized chimera antibody light chain expression vector in which said cDNA coding for the light chain constant region of human antibody and said cDNA coding for the light chain variable region of nonhuman animal antibody are linked together through said synthetic DNA; (7) introducing these expression vectors into host cells to obtain a transformant; and (8) culturing said transformant in an appropriate culture medium, thereby allowing the transformant to produce and accumulate a humanized chimera antibody, and collecting said humanized chimera antibody from the resulting culture broth.
Owner:KYOWA HAKKO KIRIN CO LTD

Enhanced first generation adenovirus vaccines expressing codon optimized HIV1-Gag, Pol, Nef and modifications

First generation adenoviral vectors and associated recombinant adenovirus-based HIV vaccines which show enhanced stability and growth properties and greater cellular-mediated immunity are described within this specification. These adenoviral vectors are utilized to generate and produce through cell culture various adenoviral-based HIV-1 vaccines which contain HIV-1 gag, HIV-1 pol and / or HIV-1 nef polynucleotide pharmaceutical products, and biologically relevant modifications thereof. These adenovirus vaccines, when directly introduced into living vertebrate tissue, preferably a mammalian host such as a human or a non-human mammal of commercial or domestic veterinary importance, express the HIV1-Gag, Pol and / or Nef protein or biologically modification thereof, inducing a cellular immune response which specifically recognizes HIV-1. The exemplified polynucleotides of the present invention are synthetic DNA molecules encoding HIV-1 Gag, encoding codon optimized HIV-1 Pol, derivatives of optimized HIV-1 Pol (including constructs wherein protease, reverse transcriptase, RNAse H and integrase activity of HIV-1 Pol is inactivated), HIV-1 Nef and derivatives of optimized HIV-1 Nef, including nef mutants which effect wild type characteristics of Nef, such as myristylation and down regulation of host CD4. The adenoviral vaccines of the present invention, when administered alone or in a combined modality regime, will offer a prophylactic advantage to previously uninfected individuals and / or provide a therapeutic effect by reducing viral load levels within an infected individual, thus prolonging the asymptomatic phase of HIV-1 infection.
Owner:EMINI EMILIO A +7

Use of cytokines secreted by dendritic cells

A prerequisite of proteomics is the ability to quantify many selected proteins simultaneously. Fluorescent sandwich immunoassays on microarrays hold appeal for such studies, since equipment and antibodies are readily available, and assays are simple, scalable and reproducible. To attain adequate sensitivity and specificity, however, a general method of immunoassay amplification is required. Coupling of isothermal rolling circle amplification (RCA) to universal antibodies can be used for this purpose: RCA on a synthetic DNA circle is initiated by a complementary oligonucleotide attached to an anti-biotin antibody; single-stranded RCA product remains attached to the antibody, and is detected by hybridization of complementary, fluorescent oligonucleotides. 51 cytokines were measured simultaneously on microarrays with signal amplification by RCA with high specificity, femtomolar sensitivity and 4 log quantitative range. This cytokine microarray was used to measure secretion from human Dendritic cells (DCs) induced by lipopolysaccharide (LPS) or tumor necrosis factor-alpha (TNF-(alpha). Rapid secretion of inflammatory cytokines such as MIP-1beta, IL-8, and IP-10 was induced by LPS. Eotaxin-2 and I-309 were found to be induced by LPS, and MDC, TARC, sIL-6R, and sTNF-RI were found to be induced by TNF-alpha. Since microarrays can accommodate ~1000 sandwich immunoassays of this type, a relatively small number of RCA microarrays appears to offer a tractable approach for proteomic surveys.
Owner:MOLECULAR STAGING

Optimized expression of HPV31 L1 in yeast

Synthetic DNA molecules encoding the HPV31 L1 protein are provided. Specifically, the present invention provides polynucleotides encoding HPV31 L1 protein, wherein said polynucleotides are free from internal transcription termination signals that are recognized by yeast. Also provided are synthetic polynucleotides encoding HPV31 L1 wherein the polynucleotides have been codon-optimized for high level expression in a yeast cell. The synthetic molecules may be used to produce HPV31 virus-like particles (VLPs), and to produce vaccines and pharmaceutical compositions comprising the HPV31 VLPs. The vaccines of the present invention provide effective immunoprophylaxis against papillomavirus infection through neutralizing antibody and cell-mediated immunityHPV31 L1 total rebuild nucleotide and aminoacid sequence:     M  S  L  W   R  P  S   1 ATGTCTTTGT GGAGACCATC E  A  T   V  Y  L  P   P  V  PTGAAGCTACC GTCTACTTGC CACCAGTCCC      V  S  K   V  V  S  T  51 AGTCTCTAAG GTCGTCTCTA  D  E  Y   V  T  R   T  N  I  YCCGACGAATA CGTCACCAGA ACCAACATCT       Y  H  A   G  S  A 101 ACTACCACGC TGGTTCTGCTR  L  L  T   V  G  H   P  Y  YAGATTGTTGA CCGTCGGTCA CCCATACTAC     S  I  P  K   S  D  N 151 TCTATCCCAA AGTCTGACAA P  K  K   I  V  V  P   K  V  SCCCAAAGAAG ATCGTCGTCC CAAAGGTCTC      G  L  Q   Y  R  V  F 201 TGGTTTGCAA TACAGAGTCT  R  V  R   L  P  D   P  N  K  FTCAGAGTCAG ATTGCCAGAC CCAAACAAGT       G  F  P   D  T  S 251 TCGGTTTCCC AGACACCTCTF  Y  N  P   E  T  Q   R  L  VTTCTACAACC CAGAAACCCA AAGATTGGTC     W  A  C  V   G  L  E 301 TGGGCTTGTG TCGGTTTGGA V  G  R   G  Q  P  L   G  V  GAGTCGGTAGA GGTCAACCAT TGGGTGTCGG      I  S  G   H  P  L  L 351 TATCTCTGGT CACCCATTGT  N  K  F   D  D  T   E  N  S  NTGAACAAGTT CGACGACACC GAAAACTCTA       R  Y  A   G  G  P 401 ACAGATACGC TGGTGGTCCAG  T  D  N   R  E  C   I  S  MGGTACCGACA ACAGAGAATG TATCTCTATG     D  Y  K  Q   T  Q  L 451 GACTACAAGC AAACCCAATT C  L  L   G  C  K  P   P  I  GGTGTTTGTTG GGTTGTAAGC CACCAATCGG      E  H  W   G  K  G  S 501 TGAACACTGG GGTAAGGGTT  P  C  S   N  N  A   I  T  P  GCTCCATGTTC TAACAACGCT ATCACCCCAG       D  C  P    P L  E 551 GTGACTGTCC ACCATTGGAAL  K  N  S   V  I  Q   D  G  DTTGAAGAACT CTGTCATCCA AGACGGTGAC     N  V  D  T   G  F  G 601 ATGGTCGACA CCGGTTTCGG A  N  D   F  T  A  L   Q  D  TTGCTATGGAC TTCACCGCTT TGCAAGACAC      K  S  W   V  P  L  D 651 CAAGTCTAAC GTCCCATTGG  I  C  N   S  I  C   K  Y  P  DACATCTGTAA CTCTATCTGT AAGTACCCAG       Y  L  K   M  V  A 701 ACTACTTGAA GATGGTCGCTE  P  Y  G   D  T  L   F  F  YGAACCATACG GCGACACCTT GTTCTTCTAC      L R  R  E   Q  M  F 751 TTGCGTAGAG AACAGATGTT V  R  H   F  F  N  R   S  G  TCGTAAGGCAC TTCTTCAACA GATCCGGCAC      V  G  E   S  V  P  T 801 CGTAGGTGAA TCTGTCCCAA  D  L  Y   I  K  G   S  G  S  TCCGACCTGTA CATCAAGGGC TCCGGTTCCA       A  T  L   A  N  S 851 CCGCTACCCT GGCTAACTCCT  Y  F  P   T  P  S   G  S  NACCTACTTCC CAACTCCATC TGGCTCCATG     V  T  S  D   A  Q  I 901 GTCACCTCCG ACGCTCAGAT F  N  K   P  Y  W  M   Q  R  ACTTCAACAAG CCATACTGGA TGCAGCGTGC      Q  G  H   N  N  G  I 951 ACAGGGTCAC AACAACGGTA  C  W  G   N  Q  L   F  V  T  VTCTGTTGGGG TAACCAGCTG TTCGTGACTG       V  D  T   T  R  S1001 TGGTCGATAC CACGCGTTCTT  N  N  S   V  C  A  A  I   AACCAACATGT CTGTCTGTGC TGCAATCGCT     N  S  D  T   T  F  K1051 AACTCTGACA CTACCTTCAA S  S  N   F  K  E  Y   L  R  HGTCCTCTAAC TTCAAGGAGT ACCTGAGACA      G  E  E   F  D  L  Q1101 TGGTGAGGAA TTCGATCTGC  F  I  F   Q  L  C   K  I  T  LAATTCATCTT CCAGTTGTGC AAGATCACCC       S  A  D   I  N  T1151 TGTCTGCTGA CATCATGACCY  I  H  S   M  N  P   A  I  LTACATCCACA GTATGAACCC TGCCATCCTG     E  D  W  N   F  G  L1201 GAGGACTGGA ACTTCGGTCT T  T  P   P  S  G  S   L  E  DGACCACTCCA CCTTCCGGTT CTTTGGAAGA.
Owner:MERCK SHARP & DOHME LLC

DNA storage method and system and storage medium

The invention discloses a DNA storage method and system and a storage medium, and the method comprises the steps of carrying out the sequencing of synthetic DNA, and obtaining a plurality of sequencing sequences; determining that no corresponding sequencing base sequence exists in the coding table, and obtaining the base sequence with the smallest Hamming distance from the sequencing base sequencein the coding table to replace the sequencing base sequence; decoding the replaced sequencing sequence according to the coding table to obtain a decoded text, wherein the Hamming distance between anytwo base sequences in the character coding table is greater than a first threshold value. According to the embodiment of the invention, error correction is carried out on the sequencing base sequenceobtained by sequencing by utilizing the Hamming distance with the base sequence, and the sequencing base sequence after error correction is decoded to obtain a decoded text. Due to the fact that theHamming distance between any two base sequences in the coding table is larger than the first threshold value, the coding table has certain error correction capacity, and compared with existing redundant code error correction, the DNA storage efficiency is greatly improved. The invention can be widely applied to the field of molecular biology.
Owner:GUANGZHOU UNIVERSITY
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