Abstract: The present invention relates to novel muteins derived from tear lipocalin or a homologue thereof. In particular, the invention relates to a mutein of human tear lipocalin. The invention also refers to a corresponding nucleic acid molecule encoding such a mutein and to a method for its generation. The invention further refers to a method for producing such a mutein. Finally, the invention is directed to a pharmaceutical composition comprising such a lipocalin mutein as well as to various use of the mutein.

Abstract: Disclosed is a method for generating a mutein of a protein selected from the group consisting of human neutrophil gelatinase-associated lipocalin (hNGAL), rat ?2-microglobulin-related protein (A2m) and mouse 24p3/uterocalin (24p3), said mutein having detectable affinity to a given target. The method comprises the step of subjecting the protein to mutagenesis at one or more of the sequence positions which correspond to the sequence positions 33 to 54, 66 to 83, 94 to 106, and 123 to 136 of hNGAL, resulting in one or more mutein(s) of the protein. Also disclosed are muteins obtainable by this method.

Abstract: The present invention relates to novel muteins derived from a bilin-binding protein (BBP) that binds a given target, for example a macromolecular target, with detectable affinity. In particular, the invention relates to a mutein of the bilin-binding protein of Pieris brassicae. The invention also refers to a corresponding nucleic acid molecule encoding such a mutein and to a method for its generation. The invention further refers to a method for producing such a mutein. Finally, the invention is directed to a pharmaceutical composition comprising such a lipocalin mutein as well as to various use of the mutein.

Abstract: A method of generating a mutein of human apolipoprotein D having detectable binding affinity to a given non-natural ligand of apolipoprotein D is disclosed, which comprises the steps of subjecting apolipoprotein D to mutagenesis at the sequence positions 34 to 38, 60, 62 to 66, 68, 89 to 93, 115, 117 to 121, and 123, resulting in a plurality of muteins of apolipoprotein D; and enriching resulting muteins having binding affinity for a given ligand from the plurality of muteins by selection, and/or isolating said mutein. Muteins of apolipoprotein D obtained by this method are also disclosed.

Abstract: The present invention refers to an isolated truncated Nogo-A polypeptide that corresponds to a truncated form of the Nogo-A protein consisting of the amino acids 174 to 940 of the full length protein of rat Nogo-A or of the amino acids 246 to 966 of the human full length Nogo-A protein.

Abstract: A method for generating a mutein of human apolipoprotein D having detectable affinity to a given non-natural ligand of apolipoprotein D is disclosed, which comprises the steps of: (a) subjecting the apolipoprotein D to mutagenesis at the sequence positions 34 to 38, 60, 62 to 66, 68, 89 to 93, 115, 117 to 121, and 123 resulting in a plurality of muteins of apolipoprotein D; and (b) enriching resulting muteins having binding affinity for a given ligand from the plurality of muteins by selection, and/or isolating said mutein. Muteins of apolipoprotein D obtainable by this method are also disclosed.

Abstract: Disclosed is a method for generating a mutein of a protein selected from the group consisting of human neutrophil gelatinase-associated lipocalin (hN-GAL), rat ?2-microglobulin-related protein (A2m) and mouse 24p3/uterocalin (24p3), said mutein having detectable affinity to a given target. The method comprises the step of subjecting the protein to mutagenesis at one or more of the sequence positions which correspond to the sequence positions 33 to 54, 66 to 83, 94 to 106, and 123 to 136 of hNGAL, resulting in one or more mutein(s) of the protein. Also disclosed are muteins obtainable by this method.

Abstract: The invention relates to the production of novel proteins exhibiting bonding activity for certain ligands, the so-called anticalins. To this end, the structure of peptides of the lipocalin family is modified by amino acid replacement in their natural ligand binding pocket using generic engineering methods. Alike immunoglobulin, the anticalin thus obtained can be used to identify or bond molecular structures.

Abstract: A method for generating a mutein of human apolipoprotein D having detectable affinity to a given non-natural ligand of apolipoprotein D is disclosed, which comprises the steps of (a) subjecting the apolipoprotein D to mutagenesis at the sequence positions 34 to 38, 60, 62 to 66, 68, 89 to 93, 115, 117 to 121, and 123 resulting in a plurality of muLeins of apolipoprutein D; and (b) enriching resulting muteins having binding affinity for a given ligand from the plurality of muteins by selection, and/or isolating said mutein. Muteins of apolipoprotein D obtainable by this method are also disclosed.

Abstract: The invention concerns a process for increasing the formation of the natural protein conformation when disulfide-bonded proteins are secreted by a prokaryotic host organism that contains a recombinant DNA coding for the secreted protein whereby the host organism is cultured in a suitable culture medium under suitable conditions for the expression of the recombinant DNA, which is characterized in that a culture medium containing 0.1 to 20 mmol/l of one or several thiol reagents is used.

Abstract: The invention concerns a polypeptide selected from muteins of streptavidin which is characterized in that it (a) contains at least one mutation in the region of the amino acid positions 44 to 53 with reference to wild type-(wt)-streptavidin and (b) has a higher binding affinity than wt-streptavidin for peptide ligands comprising the amino acid sequence Trp-X-His-Pro-Gln-Phe-Y-Z in which X represents an arbitrary amino acid and Y and Z either both denote Gly or Y denotes Glu and Z denotes Arg or Lys. In addition nucleic acids coding for the polypeptide, a vector containing this nucleic acid, a cell transfected with the vector as well as the use of a polypeptide in a method for the isolation, purification or determination of proteins are disclosed. Yet a further subject matter is a reagent kit containing the polypeptide.

Abstract: The present invention concerns a prokaryotic vector which contains a regulatable expression control sequence that can be repressed by the repressor of the tetracycline resistance gene, a prokaryotic cell transformed with this vector and the use of the vector or the cell in a process for the production of polypeptides in prokaryotes by genetic engineering.

Abstract: A peptide according to the invention comprises the amino acid sequenceTrp--X--His--Pro--Gln--Phe--Y--Z,in which X represents any desired amino acid and Y and Z either both denote Gly, or Y denotes Glu and Z denotes Arg or Lys. A fusion protein according to the invention consists of the amino acid sequence of a complete protein, of a protein mutant such as a deletion mutant or substitution mutant, or of part of a protein linked to the sequence of a peptide of the invention. For the production of a recombinant protein by expression of a DNA sequence coding therefor in suitable host cells according to well-known methods, a DNA sequence is used which codes for a fusion protein and, if desired, the presence of the expression product is detected by means of a conjugate of streptavidin and a label or the desired protein is separated as a fusion protein by means of streptavidin affinity chromatography.