tRNA synthetase fragments
View Patent ↗The present invention relates to compositions and methods for treating conditions associated with angiogenesis. In particular the present invention relates to multi-unit complexes of tRNA synthetase fragments and uses thereof; diverse multi-unit complexes including a tRNA synthetase fragment; compositions and methods for modulating angiogenesis; polynucleotides encoding tRNA synthetase fragments and uses thereof; antibodies and epitopes specific to tRNA synthetase fragments; variants of tRNA synthetase fragments and uses thereof; methods for treating angiogenesis; methods for screening for anti-angiogenic agents; methods of modulating angiogenesis; kits for modulating angiogenesis; and business methods for modulating angiogenesis. Preferably the tRNA synthetase fragments are tryptophanyl tRNA synthetase fragments, and more preferably human tryptophanyl tRNA synthetase fragments.
1. A method for purifying a multiunit complex comprising a first tryptophanyl tRNA synthetase fragment and a second tryptophanyl tRNA synthetase fragment, wherein the purified complex comprises less than 10 endotoxin units per milligram of synthetase fragment, and wherein the method comprises performing an endotoxin-reduction filtration step after performing a clarification step and prior to performing at least one of the steps selected from: a buffer exchange; a concentration step; and a cation-exchange chromatographic step, wherein said endotoxin-reduction filtration step does not include the use of a detergent.
2. A method for purifying a multiunit complex comprising a first tryptophanyl tRNA synthetase fragment and a second tryptophanyl tRNA synthetase fragment, wherein the purified complex comprises less than 10 endotoxin units per milligram of synthetase fragment, and wherein the method comprises performing an endotoxin-reduction filtration step after performing an anion-exchange chromatographic step, wherein said endotoxin-reduction filtration step does not include the use of a detergent.
3. The method according to claim 1 , wherein said first tRNA synthetase fragment and said second tRNA synthetase fragments are identical.
4. The method according to claim 3 , wherein said first tRNA synthetase fragment consists of SEQ ID NO: 27.
5. The method according to claim 1 , wherein said first tRNA synthetase fragment consists of SEQ ID NO: 27 and said second tRNA synthetase fragment consists of SEQ ID NO: 24 or SEQ ID NO: 27.
6. The method according to claim 1 , wherein said first tRNA synthetase fragment and said second tRNA synthetase fragment are each independently selected from the group consisting of SEQ ID Nos. 24, SEQ ID No. 25, SEQ ID No. 26, SEQ ID No. 27, SEQ ID No. 28, SEQ ID No. 29, SEQ ID No. 36, SEQ ID No. 37, SEQ ID No. 38, SEQ ID No. 39, SEQ ID No. 40, SEQ ID No. 41, SEQ ID No. 48, SEQ ID No. 49, SEQ ID No. 50, SEQ ID No. 51, SEQ ID No. 52 and SEQ ID No. 53, or homologs thereof, and wherein said first and second fragments are covalently dimerized.
7. The method according to claim 1 , wherein said multiunit complex has greater than 50 angiostatin activity unites; and at least 99% level of purity.