| 1. |
- Binz, Hans, et al.
(författare)
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Method for enhancing the immunogenicity of an immunogenic compound or hapten, and use thereof for preparing vaccines
- 1994
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Patent (populärvet., debatt m.m.)abstract
- The present invention relates to a process for improving the immunogenicity of an immunogen, an antigen or a hapten, when it is administered to a host, independently of the mode of administration, characterized in that the said antigen or hapten is coupled covalently to a support molecule in order to form a complex, and in that this support molecule is a polypeptide fragment which is able to bind specifically to mammalian serum albumin. The invention also relates to the use, as a medicament, of the product which can be obtained in this way.
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| 2. |
- Binz, Hans, et al.
(författare)
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Respiratory syncytial virus protein g expressed on bacterial membrane
- 1994
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Patent (populärvet., debatt m.m.)abstract
- A method for preparing a peptide or protein, wherein (a) a DNA sequence coding for a heterologous polypeptide on a peptide sequence between amino acid residues 130 and 230 of respiratory syncytial virus protein G, sub-groups A and B, or a peptide sequence at least 80 % homologous thereto, and (b) means enabling the expression of the polypeptide on the bacterial membrane surface, are inserted into a bacterium which is not pathogenic for mammals. The resulting conjugate polypeptide and a live bacterium expressing same, pharmaceutical compositions containing them and their use for preparing a vaccine, as well as a DNA sequence coding for said polypeptide, are also disclosed.
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| 3. |
- Hober, Sophia, et al.
(författare)
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Folding of insulin-like growth factor I is thermodynamically controlled by insulin-like growth factor binding protein.
- 1994
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Ingår i: Biochemistry. - : American Chemical Society (ACS). - 0006-2960 .- 1520-4995. ; 33:22
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Tidskriftsartikel (refereegranskat)abstract
- Insulin-like growth factor I (IGF-I) is thermodynamically unable to quantitatively form its native disulfides under reversible redox conditions in vitro [Hober et al. (1992) Biochemistry 31, 1749-1756]. These results prompted the question of how IGF-I may overcome this energetic problem in its folding in vivo. Here, we report that an IGF-I precursor, IGF-I-Ea, shows disulfide-exchange folding properties similar to those of mature IGF-I and, thus, is concluded not to overcome the identified folding problem of mature IGF-I. However, correct disulfide bonds are formed very efficiently when insulin-like growth factor binding protein 1 is added in equimolar amounts to IGF-I to the refolding mixture. On the basis of these results, we propose that one important function of at least one of the six homologous insulin-like growth factor binding proteins is to assist in the formation and maintenance of the native disulfides of IGF-I. To our knowledge, this is the first example where the folding of a mammalian protein or peptide in circulation has been demonstrated to be thermodynamically controlled by its binding protein. Speculatively, this could provide a mechanism to regulate the half-life of IGF-I in vivo by altering the interaction with insulin-like growth factor binding proteins.
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| 4. |
- Holmberg, A., et al.
(författare)
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Automatic Preparation of DNA Templates for Sequencing on the ABI Catalyst Robotic Workstation
- 1994
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Ingår i: Automated DNA Sequencing and Analysis. - : Elsevier Ltd. ; , s. 139-145
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Bokkapitel (övrigt vetenskapligt/konstnärligt)abstract
- The need for automation of the various steps in DNA sequencing increases as the number of applications and genomic sequencing applications grow. It is therefore necessary to design robotic workstations for template preparation and sequencing reactions in order to allow high throughput of samples with reproducible results of high quality. Also, it is not economically feasible to use manual labor in such large-scale projects.Several protocols based on paramagnetic microbeads for the recovery and purification of DNA templates to be used for sequencing have recently been described. These protocols involve both purification of polymerase chain reaction (PCR) products (Hultman et al., 1989) and M13 templates (Fry et al., 1992) and allow the isolation of specific single-stranded DNA templates. Protocols using both T7 DNA polymerase (Tabor & Richardson, 1989) and Taq cycle sequencing (Carothers et al., 1989) have been investigated. The fact that pure single-stranded templates are obtained allows accurate sequencing of heterozygotes using T7 DNA polymerase (Gibbs et al., 1989; Syvänen et al. 1991), mixed viral populations such as human immunodeficiency virus (HIV) (Wahlberg et al., 1992a) and direct mitochondrial sequencing of hair shafts and semen (Hopgood et al., 1992). The advantages of using magnetic beads as solid support include the possibility to perform manipulations such as strand melting and hybridization in a small volume with rapid kinetics (Uhlén, 1989). It is also important to point out that the magnetic beads can be transferred with a simple pipette tool, thus allowing immobilized DNA to be aliquoted and mixed within different wells or tubes within a robotic workstation. Thus, easy liquid handling is combined with rapid phase separation using a magnetic field. Recently, several automated protocols for solid phase sequencing have been described based on the Beckman Biomek-1000 laboratory workstation (Hultman et al., 1991; Wahlberg et al., 1992b).Here, we describe an experimental model of a robotic workstation (ABI Catalyst) to allow for magnetic preparation of PCR products and/or M13 templates to be used directly for sequencing either with T7 DNA polymerase or Taq DNA polymerase. A novel user interface to allow for modifications of the protocols is also described. Thus, automatic protocols with flexible parameter settings are obtained, allowing an easy setup of integrated protocols for template preparations and sequencing reactions.
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| 5. |
- Lundeberg, Joakim, 1963-, et al.
(författare)
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Chemical method for the analysis of DNA sequences
- 1993
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Patent (populärvet., debatt m.m.)abstract
- The invention provides a method of identification of the base in a target position in a DNA sequence wherein sample DNA is subjected to amplification; the amplified DNA is immobilised and then subjected to strand separation, the non-immobilised strand being removed and an extension primer, which hybridises to the immobilised DNA immediately adjacent to the target position, is provided; each of four aliquots of the immobilised single stranded DNA is then subjected to a polymerase reaction in the presence of a dideoxynucleotide, each aliquot using a different dideoxynucleotide whereby only the dideoxynucleotide whereby only the dideoxynucleotide complementary to the base in the target position becomes incorpored; the four aliquots are then subjected to extension in the presence of all four deoxynucleotides, whereby in each aliquot the DNA which has not reacted with the dideoxynucleotide is extended to form double stranded DNA while the dideoxy-blocked DNA remains as non-extended DNA; followed by identification of the double stranded and/or non-extended DNA to indicate which dideoxynucleotide was incorporated and hence which base was present in the target position.
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| 6. |
- Nilsson, Björn, et al.
(författare)
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Bacterial receptor structures
- 1994
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Patent (populärvet., debatt m.m.)abstract
- Novel proteins obtainable by mutagenesis of surface-exposed amino acids of domains of natural bacterial receptors, said proteins being obtained without substantial loss of basic structure and stability of said natural bacterial receptors; proteins which have been selected from a protein library embodying a repertoire of said novel proteins; and methods for the manufacture of artificial bacterial receptor structures.
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| 7. |
- Strandberg, L, et al.
(författare)
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Expression and characterization of a tripartite fusion protein consisting of chimeric IgG-binding receptors and beta-galactosidase
- 1990
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Ingår i: Journal of Biotechnology. - : Elsevier. - 0168-1656 .- 1873-4863. ; 13:1, s. 83-96
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Tidskriftsartikel (refereegranskat)abstract
- Using protein engineering, a tripartite fusion protein was constructed consisting of five IgG-binding regions of protein A from Staphylococcus aureus, two IgG-binding regions of protein G from Streptococcus strain G148 and beta-galactosidase from Escherichia coli. The resulting protein lacks the serum albumin binding regions of native protein G. The fusion protein, which is a tetramer of approximately 660 kDa, was designed as a tool for immunological assays taking advantage of its broad spectrum of antibody affinity. The gene was placed under control of two promoters, the PR promoter and the lac UV5 promoter and the expression from the two promoters was studied in a bioreactor. Induction of the PR promoter gave an intracellular product concentration corresponding to 20% of the cell dry weight. By utilizing the properties of beta-galactosidase, the protein was purified by extraction in an aqueous two-phase system. The fusion protein was not proteolytically degraded during the cultivation and purification steps. The biological activity of all three parts of the protein was demonstrated with a competitive ELISA.
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| 8. |
- Ståhl, Stefan, et al.
(författare)
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Recombinant DNA coding for signal peptide, selective interacting polypeptide and membrane anchoring sequence
- 1991
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Patent (populärvet., debatt m.m.)abstract
- Tripartite recombinant DNA encoding fusion proteins which comprise three sequences, i.e., a signal peptide which is operable in a Gram positive bacterium, an immunogenic polypeptide linked thereto which is not normally expressed in a Gram positive bacterium, and a cell wall spanning and a membrane anchoring sequence, as well as their use in Gram positive bacteria to express the resultant fusion protein on their surface are described. The preferred cell wall spanning and anchoring polypeptides include Staphylococcus protein A and Streptococcus protein G.
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